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1

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

EPA Science Inventory

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

2

Potential problems with fluorescein diacetate assays of cell viability when testing natural products for antimicrobial activity  

Microsoft Academic Search

There are two potential problems in the use of fluorescein diacetate (FDA) as a measure of cell viability. The first is the hydrolysis of FDA to fluorescein in the absence of live cells and the second is the quenching of fluorescence by assay solutions. We show that common media components such as tryptone, peptone and yeast extract all promote hydrolysis

Joanne M. Clarke; Michael R. Gillings; Nanda Altavilla; Andrew J. Beattie

2001-01-01

3

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

NASA Astrophysics Data System (ADS)

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.

Yamada, Kengo; Yoshida, Katsuyuki; Sonoda, Ryoichi

4

Assessment of fluorescein-based fluorescent dyes for tracing Neotyphodium endophytes in planta.  

PubMed

Fluorescent dyes were assessed for their ability to stain viable hyphae of the fungi Neotyphodium lolii and N. coenophialum, symbiotic endophytes of the Pooideae grasses Lolium perenne and Festuca arundinacea, respectively. The fluorescein-based fluorophores; fluorescein diacetate (FDA), 5(6)-carboxy-fluorescein diacetate (CFDA), 5-chloromethylfluorescein diacetate (CMFDA) and the chitin-binding stain, Calcofluor while M2R, were assessed for staining of endophyte hyphae in vitro from axenic fungal cultures and in planta, including epidermal leaf sheath peels, nodes, ovaries, embryos and meristems. CMFDA produced the greatest intensity of staining of fungal hyphae and gave excellent contrast in planta compared to the plant cells. Compared to the other dyes, CMFDA was also the least affected by photo bleaching and continued to fluoresce up to 2 h after initial excitation. None of the fluorescent dyes stained fungal hyphae in seed. PMID:22802389

Card, Stuart D; Tapper, Brian A; Lloyd-West, Catherine; Wright, Kathryn M

2013-01-01

5

Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay: a quantitative method for oxidative stress assessment of nanoparticle-treated cells.  

PubMed

No consensus exists on how to address possible toxicity of nanomaterials as they interfere with most in vitro screening tests based on colorimetric and fluorimetric probes such as the dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay for detection of oxidative species. In the present research, nanomaterial interaction with DCFH-DA was studied in relation to its nature and/or assay conditions (cell-based and time exposure) by incubating Rhodamine (Rhd)-labeled 25nm and 50nm silica (SiO2), naked and oleic acid coated magnetite, (Fe3O4) and maghemite (Fe2O3) iron oxide, titanium dioxide (TiO2) and poly(ethylene oxide)-poly(lactide/glycolide) acid (PLGA-PEO) nanoparticles (NPs) with metabolically active rat hepatocytes for 4 and 24-h periods. Data indicated that nanoparticle uptake correlated with quenching of dye fluorescence emission. In spite of their masking effect, the oxidative potential of NPs could be detected at a limited threshold concentration when exposed for periods of time longer than those frequently used for this test. However, changes in the experimental conditions did not systematically result in free radical formation for all nanomaterials tested. Overall data indicate that despite the quenching effect of nanoparticles on DCFH-DA assay, it can be considered as a useful tool for quantitative measurement of NPs-induced oxidative stress by minor modifications of standardized protocols. PMID:23357416

Aranda, A; Sequedo, L; Tolosa, L; Quintas, G; Burello, E; Castell, J V; Gombau, L

2013-03-01

6

Use of fluorescent staining and flow cytometry for monitoring physiological changes in solventogenic clostridia.  

PubMed

Physiological changes in populations of Clostridium beijerinckii and Clostridium tetanomorphum were monitored by fluorescence staining and flow cytometry. To estimate the number of metabolically active cells in exponential growth, a combination of the dyes propidium iodide and carboxy fluorescein diacetate appeared to be a good choice for both species. During stationary phase, these stains did not reflect physiological changes sufficiently and therefore additional labeling with bis-(1,3-dibutylbarbituric acid) trimethineoxonol was applied. Results of fluorescence staining in solventogenic batch fermentations were compared with substrate-use data, the concentration of key metabolites and growth curves. We demonstrate that measurements by all methods were mutually compatible. PMID:24211310

Patakova, Petra; Linhova, Michaela; Vykydalova, Pavla; Branska, Barbora; Rychtera, Mojmir; Melzoch, Karel

2014-10-01

7

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

SciTech Connect

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.

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

1995-12-31

8

The photography of fluorescein  

SciTech Connect

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

Welch, J.D.

1982-06-01

9

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

PubMed Central

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

2014-01-01

10

Measurement of retinal blood flow with fluorescein leucocyte angiography using a scanning laser ophthalmoscope in rabbits  

Microsoft Academic Search

AIMS: To measure blood flow in the rabbit retinal circulation with fluorescein leucocyte angiography using a scanning laser ophthalmoscope. METHODS: Blood was withdrawn from the ear vein of a rabbit (New Zealand White), mixed with fluorescent dye in a test tube and centrifuged. The yellow-brown layer containing fluorescein stained leucocytes was collected and injected into the ear vein of the

Y Yang; S Moon; S Lee; J Kim

1996-01-01

11

21 CFR 522.1078 - Gonadorelin diacetate tetrahydrate.  

Code of Federal Regulations, 2010 CFR

... 2010-04-01 false Gonadorelin diacetate tetrahydrate. 522.1078 Section...ANIMAL DRUGS § 522.1078 Gonadorelin diacetate tetrahydrate. (a) Specifications...contains 50 micrograms (µg) of gonadorelin diacetate tetrahydrate. (b) Sponsors...

2010-04-01

12

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.

13

Effect of novaluron on microbial biomass, respiration, and fluorescein diacetate-hydrolyzing activity in tropical soils  

Microsoft Academic Search

The aim of this study was to assess the potential harmful effects of novaluron on soil microbiological parameters in clay\\u000a loam alluvial soil (Typic udifluvent) and coastal saline soil (Typic endoaquept) under controlled laboratory tests. The applications\\u000a of novaluron were made at or above the recommended rates, which includes field rate (FR), two times (2FR), and ten times (10FR)\\u000a the

Piw Das; Raktim Pal; Ashim Chowdhury

2007-01-01

14

Stain Reagent Reversible Stain Kits  

E-print Network

are not visible before destain- ing with methanol/acetic acid to remove the characteristic background stainingGelCode ® Blue Stain Reagent GelCode ® SilverSNAP TM Stain GelCode ® E-Zinc TM Reversible Stain Kits Technical Review Gel Stains F E A T U R I N G . . . #12;P r o t e i n S t a i n i n g Only

Lebendiker, Mario

15

Benign Adenoma of the Iris Pigment Epithelium: Clinical and Iris Fluorescein Angiographic Features  

Microsoft Academic Search

Benign adenoma of iris pigment epithelium is a rare neoformation characterized by a multinodular, dark brown to dark black, relatively stationary lesion, especially localized in the peripheral iris. We report a case of iris pigment epithelium adenoma with no evidence of change during the 6-year follow-up. Iris fluorescein angiography showed a mild staining of the lesion without evidence of abnormal

V. Isola; M. Battaglia Parodi; S. Calderini

1994-01-01

16

ELECTRON STAINS  

PubMed Central

Chemical studies have been carried out on the interaction of DNA with uranyl salts. The effect of variations in pH, salt concentration, and structural integrity of the DNA on the stoichiometry of the salt-substrate complex have been investigated. At pH 3.5 DNA interacts with uranyl ions in low concentration yielding a substrate metal ion complex with a UO2++/P mole ratio of about ½ and having a large association constant. At low pH's (about 2.3) the mole ratio decreases to about ?. Destruction of the structural integrity of the DNA by heating in HCHO solutions leads to a similar drop in the amount of metal ion bound. Raising the pH above 3.5 leads to an apparent increase in binding as does increasing the concentration of the salt solution. This additional binding has a lower association constant. Under similar conditions DNA binds about seven times more uranyl ion than bovine serum albumin, indicating useful selectivity in staining for electron microscopy. PMID:13788706

Zobel, C. Richard; Beer, Michael

1961-01-01

17

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

Code of Federal Regulations, 2010 CFR

...Tolerances § 180.1058 Sodium diacetate; exemption from the requirement of a tolerance. Sodium diacetate, when used postharvest as a fungicide, is exempt from the requirement of a tolerance for residues in or on alfalfa, hay; Bermudagrass,...

2010-07-01

18

Superacidic cyclization of ?-hydroxygeraniol diacetate and ?-hydroxygeraniol benzyl ether acetate  

Microsoft Academic Search

Low-temperature superacidic cyclization of (E,E)-3,7-dimethylocta-2,6-diene-1,8-diol (?-hydroxygeraniol) diacetate and (E,E)-8-acetoxy-1-benzyloxy-3,7-dimethylocta-2,6-diene leads to the same mixtures of two diastereomeric 9-acetoxy-8-hydroxy-p-menth-1-enes epimeric at the C(8) atom.

V. Kulci?ki; N. Ungur; C. Deleanu; P. F. Vlaa

1999-01-01

19

Coffee stain on textiles. Mechanisms of staining and stain removal  

Microsoft Academic Search

Coffee stains on textiles are mainly caused by the water-soluble and acidic colored substances in coffee. The acidic nature\\u000a of coffee stain has been shown by ultraviolet and visible spectroscopy of coffee as a function of pH; ion-pair formation with\\u000a a cationic surfactant and titration with Hyamine 1622 and a surfactant-specific electrode; and precipitation of the colored\\u000a components in coffee

Erik Kissa

1995-01-01

20

Peripapillary fluorescein angiographic findings in primary open angle glaucoma  

Microsoft Academic Search

BACKGROUND: Detailed fluorescein angiographic findings in the disc circumference may be useful for evaluating the possible relation of the circumference to glaucomatous nerve damage. METHODS: Fluorescein angiograms of 25 eyes of 25 subjects with primary open angle glaucoma were observed after they had undertaken Octopus perimetry. Based on the retinotopic projection, disc sectors and corresponding visual field regions were set.

S Yamazaki; Y Inoue; K Yoshikawa

1996-01-01

21

Cryopreservation of equine oocytes using glycerol as the cryoprotectant and cumulus investment as a parameter  

E-print Network

cumulus layer; and 3) corona radiata only. All oocytes were stained with fluorescein diacetate (FDA) to determine viability before freezing. Forty viable oocytes from each group were frozen to be evaluated post-thaw using FDA and orcein stains. None...

Lippert, Jennifer Johnson

2012-06-07

22

Causes of Bridge Pier Staining.  

National Technical Information Service (NTIS)

Four types of bridge stains exist in Arkansas: Rust stains - those stains directly traceable to rust; red stains - broad stains which are not directly traceable to rust; gray stains - similar to red stains except for color; and graffiti. Bridge stains in ...

S. I. Thornton, C. Springer

1974-01-01

23

Port-wine stain  

MedlinePLUS

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

24

Ultra-widefield fluorescein angiography of white without pressure  

PubMed Central

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

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

2013-01-01

25

Acute Toxicity of Sodium Fluorescein to Ashy Pebblesnails Fluminicola fuscus  

USGS Publications Warehouse

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

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

2011-01-01

26

Fluress, fluorescein and benoxinate: recovery from bacterial contamination.  

PubMed

The ability to recover from bacterial contaminations with Staphylococcus auresus and Pseudomonas aeruginosa was determined for three optometric DPAs: Fluress, fluorescein and benoxinate. Results show that Fluress recovers from contamination more rapidly than benoxinate or fluorescein. This ability to recover from contamination and the relative ease of use of Fluress may make it the DPA choice for a number of optometric procedures including applanation tonometry. PMID:6771320

Yolton, D P; German, C J

1980-05-01

27

Understanding Romanowsky staining  

Microsoft Academic Search

Normal blood smears were stained by the standardised azure B-eosin Y Romanowsky procedure recently introduced by the ICSH, and the classical picture resulted. The effects of varying the times and temperature of staining, the composition of the solvent (buffer concentration, methanol content, & pH), the concentration of the dyes, and the mode of fixation were studied. The results are best

R. W. Horobin; K. J. Walter

1987-01-01

28

FTIR evolved gas analysis of the decomposition products of cellulose diacetate  

Microsoft Academic Search

Evolved gas analysis of unplasticised cellulose diacetate flake and plasticised cellulose diacetate film (Clarifoil) was carried out as part of an assessment of the potential hazard that could occur as a result of overheating these materials during processing. In order to perform this work, a simple apparatus for heating a sample and introducing the evolved gases into an FTIR gas

D. M. Price; S. P. Church

1997-01-01

29

Gram stain of tissue biopsy  

MedlinePLUS

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

30

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

E-print Network

. Meaty/brothy complex, smoke, spice aroma, springiness, and cohesiveness attributes were judged slightly lower for frankfurters formulated without lactate/diacetate than those with lactate/diacetate at the end of aerobic storage. Sensory color...

Knight, Timothy David

2006-08-16

31

Localization of Legionella pneumophila in tissue using FITC-conjugated specific antibody and a background stain  

SciTech Connect

Lightly staining formalin-fixed or fresh tissue with Gram's crystal violet obviates interfering nonspecific fluorescence by acting as a metachromatic stain in ultraviolet light. Against the easily recognized background of tissues and cells fluorescein isothiocyanate-tagged Legionella pneumophila antibodies can then identify this bacterium in or on individual cells. This procedure can be run at room temperature in two hours and has the potential for further widespread applicability.

Lowry, B.S.; Vega, F.G. Jr.; Hedlund, K.W.

1982-05-01

32

Pericardial fluid Gram stain  

MedlinePLUS

... staining a sample of fluid taken from the sac surrounding the heart to diagnose a bacterial infection. ... sample of fluid will be taken from the sac surrounding the heart. Before this is done, some ...

33

Quick Stain Removal Guide  

E-print Network

paints) rinse garment in warm water. For oil-based paints, apply solvent recommended on paint can or use turpentine. Rinse. Pretreat with prewash stain remover, bar soap or laundry detergent. Rinse and wash. Acne medicine Adhesive tape, chewing gum... paints) rinse garment in warm water. For oil-based paints, apply solvent recommended on paint can or use turpentine. Rinse. Pretreat with prewash stain remover, bar soap or laundry detergent. Rinse and wash. Acne medicine Adhesive tape, chewing gum...

Brown, Pamela J.

1998-07-29

34

Purification of murine endothelial cell cultures by flow cytometry using fluorescein-labeled griffonia simplicifolia agglutinin.  

PubMed Central

Griffonia simplicifolia agglutinin (GSA) is a valuable histochemical tool in the identification of endothelium. In this study GSA labeled with fluorescein isothiocyanate (GSA-FITC) was used to purify cultures of murine cerebral microvascular endothelium. Cultures were stained with GSA-FITC, then sorted using a fluorescence-activated cell sorter (FACS). GSA-positive endothelial cells were collected, re-cultured, and subsequently re-analyzed by FACS using GSA-FITC. Cultures that initially contained 80 +/- 3 to 89 +/- 3% (X +/- SE) GSA-positive cells were purified to 98 +/- 1% positivity. Immunohistochemistry with an anti-muscle-action antibody confirmed that FACS sorting of GSA-FITC-stained cells effectively removed contaminating smooth muscle cells from endothelial cell cultures. Viability, proliferation, and prostaglandin production of the cells was unaltered by lectin staining and FACS sorting. Thus, GSA-FITC can be used in conjunction with flow cytometry to enhance the purity of murine endothelial cell cultures without adversely affecting cell viability, growth, or metabolism. Images Figure 2 PMID:2757116

Sahagun, G.; Moore, S. A.; Fabry, Z.; Schelper, R. L.; Hart, M. N.

1989-01-01

35

Effects of topical anaesthetics and fluorescein on the real-time PCR used for the diagnosis of Herpesviruses and Acanthamoeba keratitis  

PubMed Central

Background The early microbiological diagnosis of corneal infections may prevent the condition from worsening. Aim To study the potential interferences of oxybuprocain and fluorescein solutions used by ophthalmologists on the performances of the real?time polymerase chain reaction (PCR) carried out as routine test for diagnosis of keratitis. Methods Quantified suspensions of Herpes simplex virus (HSV1), Varicella zoster virus (VZV), Cytomegalovirus (CMV) and Acanthamoeba with and without oxybuprocain or fluorescein added before DNA extraction were tested by real?time PCR. Results The capacities of the real?time PCR to detect HSV, VZV, CMV and Acanthamoeba were reduced by oxybuprocain and fluorescein. Both products diluted to 1/16 reduced the PCR detection capacities for more than 2?logs (DNA copies/sample). Conclusions The simultaneous introduction of fluorescein or topical anaesthetics into the tubes containing the specimens to be tested by PCR may lead to false negative results. Because corneal specimens for microbiological diagnosis of keratitis are obtained after topical administration of anaesthetics and corneal staining with fluorescein, ophthalmologists should be aware to rinse the eye surface intensively with appropriate eye solutions to minimise the risks of misdiagnosis. PMID:16899529

Goldschmidt, P; Rostane, H; Saint-Jean, C; Batellier, L; Alouch, C; Zito, E; Bourcier, T; Laroche, L; Chaumeil, C

2006-01-01

36

Kinetics of staining and bleaching  

Microsoft Academic Search

Kinetics of staining and stain removal have been studied with food dyes and natural colorants. Kinetics of staining indicate\\u000a that the staining agent is adsorbed on the fiber surface and diffuses into the interior of the fibers. Similarities exist\\u000a between staining and dyeing mechanisms of cotton, polyester and nylon fibers. Stain removal by nonoxidative detergency involves\\u000a diffusion of the staining

Erik Kissa; Jenny M. Dohner; Ward R. Gibson; Donna Strickman

1991-01-01

37

Digital Angiography Of The Ocular Fundus Without Fluorescein  

NASA Astrophysics Data System (ADS)

Local spatial filters were developed to enhance and extract blood vessels in a digitized color non-fluorescein angiograph. One filter dynamically enhances a black/white digital picture by normalizing the pixels relative to their local neighborhoods, and another filter extracts the candidate blood vessel pixels from a black/white digital picture.

Starr, Lee J.; Ishaq, Naseem

1988-06-01

38

A mitochondria-targeted protonophoric uncoupler derived from fluorescein.  

PubMed

Linking decyl-triphenyl-phosphonium to fluorescein yields a fluorescent probe that accumulates in energized mitochondria, facilitates proton transfer across membranes and stimulates mitochondrial respiration. This features a mitochondria-targeted uncoupler, being of potential interest for therapeutic use against oxidative stress-related diseases. PMID:25349923

Denisov, Stepan S; Kotova, Elena A; Plotnikov, Egor Y; Tikhonov, Artur A; Zorov, Dmitry B; Korshunova, Galina A; Antonenko, Yuri N

2014-12-18

39

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.

40

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

PubMed

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

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

2014-01-01

41

Image Acquisition And Storage For Ophthalmic Fluorescein Angiography  

NASA Astrophysics Data System (ADS)

An ophthalmic imaging system, the IS-2000, has been developed for acquisition, archival storage, and analysis of fundus imagery collected during fluorescein angiography and other ophthalmic procedures. The system consists of a conventional fundus camera equipped with a video camera; a microcomputer with color video digitizer and frame buffer; and an optical video disk for archival storage of images. In the course of development of the IS-2000 a study was performed to assess the spatial frequency content of typical angiograms and to evaluate the resolution capabilities of a variety of types of commercially available video cameras and image storage media. The frequency content of angiograms was analyzed through the application of Fourier transforms to high-resolution digitized images. Modulation transfer functions (MTF) of various types of video cameras and video disks were measured with appropriate video test images. An assessment of the suitability of currently available video equipment for collection and storage of fluorescein angiograms is provided.

Cambier, J. L.; Nelson, M. R.; Brown, S. I.; Goldbaum, M. H.; Rehkopf, P. G.; Warnicki, J. L.

1984-08-01

42

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...

43

Length of stain dosimeter  

NASA Technical Reports Server (NTRS)

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.

Lueck, Dale E. (inventor)

1994-01-01

44

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

SciTech Connect

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.

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

45

Gram stain of skin lesion  

MedlinePLUS

Skin lesion gram stain ... a glass slide. A series of different colored stains is applied to the sample. A laboratory team ... test. For information on risks related to the removal of a skin sample, see skin lesion biopsy .

46

Removing Stains from Washable Fabrics.  

E-print Network

protein and dye. d. Special stains needing unique treatment be cause of chemical make-up or physical characteristics. (Examples: chewing gum, iodine, lead pencil) Stain Removal Products Bleaches Chlorine bleaches contain a hypochlorite com pound... Beer Combination 8 Wet 8 Protein 8 Tannin 8 Chewing gum Benzoil peroxide Special 9 Special 9 Chocolate Blood Combination 8 Wet 8 Dye 8 Protein 8 4 Stain Page Numbers Stain Page Numbers Cocoa Feces Combination 8 Wet 8 Dye 8 Protein 8 Coffee...

Beard, Ann Vanderpoorten

1988-01-01

47

Digitization of stained glass  

NASA Astrophysics Data System (ADS)

Digital photography was applied to the capture of images of the stained glass windows in the historic parish church in Fairford, Gloucestershire, England. Because of their size, the windows had to be photographed in 45 separate sections in order to capture all the detail present in the painting on the glass. The digital images of each section, approximately 3000 by 2300 pixels, were then mosaiced together in order to construct the very high resolution image needed for the complete window. A special backlight panel was constructed for the purpose, and techniques developed for minimizing the effects of reflected light and for calibrating the color of the images. Improvements in the technology for mounting and positioning the camera were identified as the most significant factors currently preventing the widespread adoption of this technology for virtual heritage applications.

MacDonald, Lindsay W.

1997-04-01

48

Susac's syndrome: the value of fundus fluorescein angiography.  

PubMed

A 19-year-old woman presented with a 4-week history of headache, ataxia, vertigo, confusion, intermittent blurred vision in the right eye and intermittent hearing loss. MRI revealed white matter lesions and 'pepper pot' lesions of the corpus callosum. The cerebrospinal fluid had raised protein and lymphocytes. Fundal examination revealed multiple peripheral arterial occlusions in the both eyes confirmed with fundus fluorescein angiography (FFA). A diagnosis of Susac's syndrome was made. The patient was initially treated with steroids, followed by azathioprine and intravenous immunoglobulins (IVIg). Clinical improvement was noted, associated with improvement of the retinal circulation on FFA. PMID:25281252

Khan, Imran Joseph; Allroggen, Holger; Pagliarini, Sergio

2014-01-01

49

Influence of a cellulose diacetate matrix on the complexation kinetics of tetraphenylporphin with Zn and Cd  

NASA Astrophysics Data System (ADS)

The dependence of the reaction rate of tetraphenylporphin zinc and cadmium complexes in a polymer matrix on a base of cellulose diacetate and low-molecular model solutions was investigated. The characteristics of the diffusive transport of aqueous solutions of zinc and cadmium acetates through the cellulose diacetate membrane were obtained. The kinetic control of the porphyrin reaction incorporated into the polymer, and the determining influence of the steric limitations of the matrix of a rigid chain polymer on macroheterocycle deformation (and thus its reactivity) are shown.

Trifonova, I. P.; Kononov, V. D.; Burmistrov, V. A.; Koifman, O. I.

2011-04-01

50

Meibomian orifices and Marx's line. Studied by triple vital staining.  

PubMed

The ciliary margins of the lower lids have been vital stained by the lipid-specific Sudan III powder, fluorescein 0.1% and the bottom of the lacrimal river (Marx's line) by lissamine green 1% in 100 cases. The Meibomian orifices are situated in a straight row just in front of the Marx's line in the lipid phase. With increasing age (greater than 50 years) the orifices are more often displaced and also discharge their lipid in the depth of the aqueous phase. The number averaged 21.5 in the lipid phase and 1.7 in the aqueous phase. Active orifices staining with lipid were found in 45% of all orifices in normals, independent of age, and were increased in conjunctivitis in the lipid phase. Lissamine green-stained orifices were independent of age, phase and diagnosis. The anterior edge of Marx's line may run an irregular course in elderly normals (greater than 50 years), significantly more often in conjunctivitis and blepharitis. PMID:2420147

Norn, M

1985-12-01

51

CNT Uptake in Human Ovarian Cancer Cells: Changes in Fluorescence Emission of CNT-Fluorescein in  

E-print Network

CNT Uptake in Human Ovarian Cancer Cells: Changes in Fluorescence Emission of CNT-Fluorescein in SKOV-3 Ovarian Cancer Cells Meng-Tse Chen 1, Lewis Gomez-De Arco 2,Yinghua Sun1, P. Thomas Vernier3-linked carbon nanotube (CNT)- fluorescein conjugates into SKOV-3 human ovarian cancer cells. The uptake of CNTs

Southern California, University of

52

The role of intravenous fluorescein in the detection of colon ischemia during aortic reconstruction  

Microsoft Academic Search

Intravenous fluorescein is an accurate predictor of small bowel viability, but its effectiveness in assessing colon perfusion during aortic surgery has not been evaluated. Over a 10 year period 186 of 3,306 patients undergoing aortic reconstruction received 500 to 1000 mg of intravenous fluorescein intraoperatively to evaluate colon viability. Prior history of colectomy, hypogastric or mesenteric arterial occlusive disease, or

R. Thomas Bergman; Peter Gloviczki; Timothy J. Welch; James M. Naessens; Thomas C. Bower; John W. Hallett; Peter C. Pairolero; Kenneth J. Cherry

1992-01-01

53

Development of a Vital Fluorescent Staining Method for Monitoring Bacterial Transport in Subsurface Environments  

PubMed Central

Previous bacterial transport studies have utilized fluorophores which have been shown to adversely affect the physiology of stained cells. This research was undertaken to identify alternative fluorescent stains that do not adversely affect the transport or viability of bacteria. Initial work was performed with a groundwater isolate, Comamonas sp. strain DA001. Potential compounds were first screened to determine staining efficiencies and adverse side effects. 5-(And 6-)-carboxyfluorescein diacetate, succinimidyl ester (CFDA/SE) efficiently stained DA001 without causing undesirable effects on cell adhesion or viability. Members of many other gram-negative and gram-positive bacterial genera were also effectively stained with CFDA/SE. More than 95% of CFDA/SE-stained Comamonas sp. strain DA001 cells incubated in artificial groundwater (under no-growth conditions) remained fluorescent for at least 28 days as determined by epifluorescent microscopy and flow cytometry. No differences in the survival and culturability of CFDA/SE-stained and unstained DA001 cells in groundwater or saturated sediment microcosms were detected. The bright, yellow-green cells were readily distinguished from autofluorescing sediment particles by epifluorescence microscopy. A high throughput method using microplate spectrofluorometry was developed, which had a detection limit of mid-105 CFDA-stained cells/ml; the detection limit for flow cytometry was on the order of 1,000 cells/ml. The results of laboratory-scale bacterial transport experiments performed with intact sediment cores and nondividing DA001 cells revealed good agreement between the aqueous cell concentrations determined by the microplate assay and those determined by other enumeration methods. This research indicates that CFDA/SE is very efficient for labeling cells for bacterial transport experiments and that it may be useful for other microbial ecology research as well. PMID:11010903

Fuller, Mark E.; Streger, Sheryl H.; Rothmel, Randi K.; Mailloux, Brian J.; Hall, James A.; Onstott, Tullis C.; Fredrickson, James K.; Balkwill, David L.; DeFlaun, Mary F.

2000-01-01

54

Spore Stain of Bacillus cereus  

NSDL National Science Digital Library

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.

American Society For Microbiology;

2002-01-01

55

Fluorescein angiography to estimate normal peripheral retinal nonperfusion in children  

PubMed Central

Purpose To estimate the normal distance from vascular termini to ora serrata in children’s eyes. Methods Clinical records and peripheral fluorescein angiography images of the ora serrata region, taken using scleral indentation and the RetCam system during examination under anesthesia, were retrospectively reviewed from 33 eyes of 31 consecutive patients with presumed normal peripheral retinal vasculature. All patients had ocular disease either only in the fellow eye or if in the study eye, to a degree judged not likely to affect peripheral retinal vascular development. Results The mean age at angiography was 3.8 years (range, 2 months to 13 years). Means of 0.9 disk diameters (DD) of nonperfusion temporally (range, 1.5 DD to 0.5 DD; SD 0.3) and 0.6 DD of nonperfusion nasally (range, 1 DD to 0.25 DD; SD 0.2) were found. Conclusions In children up to age 13 years, the avascular retina normally extends 1.5 DD or less temporally and 1.0 DD or less nasally from the ora serrata. Conservatively, ?2 DD of nonperfusion, 3 standard deviations more than normal, should be considered abnormal and a sign of peripheral nonperfusion. These data may serve as preliminary indicators of the range of normal when evaluating diseases with retinal vascular abnormalities in children. PMID:22681939

Blair, Michael P.; Shapiro, Michael J.; Hartnett, M. Elizabeth

2013-01-01

56

Fluorescein as a model molecular calculator with reset capability  

NASA Astrophysics Data System (ADS)

The evolution of molecules capable of performing boolean operations has gone a long way since the inception of the first molecular AND logic gate, followed by other logic functions, such as XOR and INHIBIT, and has reached the stage where these tiny processors execute arithmetic calculations. Molecular logic gates that process a variety of chemical inputs can now be loaded with arrays of logic functions, enabling even a single molecular species to execute distinct algebraic operations: addition and subtraction. However, unlike electronic or optical signals, the accumulation of chemical inputs prevents chemical arithmetic systems from resetting. Consequently, a set of solutions is required to complete even the simplest arithmetic cycle. It has been suggested that these limitations can be overcome by washing off the input signals from solid supports. An alternative approach, which does not require solvent exchange or incorporation of bulk surfaces, is to reset the arithmetic system chemically. Ultimately, this is how some biological systems regenerate. Here we report a highly efficient and exceptionally simple molecular arithmetic system based on a plain fluorescein dye, capable of performing a full scale of elementary addition and subtraction algebraic operations. This system can be reset following each separate arithmetic step. The ability to selectively eradicate chemical inputs brings us closer to the realization of chemical computation.

Margulies, David; Melman, Galina; Shanzer, Abraham

2005-10-01

57

Ultrawide-field Fluorescein Angiography for Evaluation of Diabetic Retinopathy  

PubMed Central

Purpose To investigate the advantages of ultrawide-field fluorescein angiography (FA) over the standard fundus examination in the evaluation of diabetic retinopathy (DR). Methods Ultrawide-field FAs were obtained in 118 eyes of 59 diabetic patients; 11 eyes with no DR, 71 eyes with nonproliferative diabetic retinopathy (NPDR), and 36 eyes with proliferative diabetic retinopathy (PDR), diagnosed by the standard method. The presence of peripheral abnormal lesions beyond the standard seven fields was examined. Results Ultrawide-field FA images demonstrated peripheral microaneurysms in six (54.5%) of 11 eyes with no DR and all eyes with moderate to severe NPDR and PDR. Peripheral retinal neovascularizations were detected in three (4.2%) of 71 eyes with NPDR and in 13 (36.1%) of 36 eyes with PDR. Peripheral vascular nonperfusion and vascular leakage were found in two-thirds of eyes with severe NPDR and PDR. Conclusions Ultrawide-field FA demonstrates peripheral lesions beyond standard fields, which can allow early detection and a close evaluation of DR. PMID:23204797

Kong, Mingui; Lee, Mee Yon

2012-01-01

58

Drop Coating Deposition Raman Spectroscopy of Fluorescein Isothiocyanate Labeled Protein  

PubMed Central

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

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

2011-01-01

59

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

PubMed

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

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

2009-08-01

60

Crystal structures and properties of two new pseudopolymorphic modifications of the glucocorticoide triamcinolone diacetate  

NASA Astrophysics Data System (ADS)

Two new solvates of triamcinolone diacetate were found in addition, to those reported previously. The acetonitrile solvate (form E) crystallizes monoclinic in space group P2 1, whereas the methylene chloride solvate (form F) crystallizes orthorhombic in space group P2 12 12 1. In all forms the triamcinolone diacetate molecules are linked by intermolecular hydrogen bonding. From this arrangement channels are formed in which the solvent molecules are embedded. Both forms were investigated by differential thermoanalysis and thermogravimetry. On heating, for each form a mass loss is observed, which is accompanied with endothermic events in the DTA curve. Mass spectroscopic investigations clearly shows that in this step the solvent molecules are emitted. In these measurements one cannot differ between desolvation and melting. If the residues formed after the first TG steps are investigated by X-ray powder diffraction, only amorphous samples are obtained. If the solvents are removed at room temperature under normal pressure or in vacuum the commercial available form of triamcinolone diacetate is obtained which is also used in therapy. If the acetonitrile solvate is tempered at 80 °C for several days significant changes in the powder pattern are observed, which may indicate the formation of a new polymorphic form.

Suitchmezian, Viktor; Jeß, Inke; Näther, Christian

2006-11-01

61

Transscleral diffusion of ethacrynic acid and sodium fluorescein  

PubMed Central

Purpose One of the current limitations in developing novel glaucoma drugs that target the trabecular meshwork (TM) is the induced corneal toxicity from eyedrop formulations. To avoid the corneal toxicity, an alternative approach would be to deliver TM drugs through the sclera. To this end, we quantified ex vivo diffusion coefficient of a potential TM drug, ethacrynic acid (ECA), and investigated mechanisms of ECA transport in the sclera. Methods An Ussing-type diffusion apparatus was built to measure the apparent diffusion coefficient of ECA in fresh porcine sclera at 4 °C. To understand mechanisms of ECA transport, we quantified the transscleral transport of a fluorescent tracer, sodium fluorescein (NaF), that has a similar molecular weight but is more hydrophilic compared to ECA. Furthermore, we developed a mathematical model to simulate the transport processes and used it to analyze the experimental data. The model was also used to investigate the dependence of diffusion coefficients on volume fraction of viable cells and the binding of NaF and ECA to scleral tissues. Results The diffusion coefficients of ECA and NaF in the sclera were 48.5±15.1x10-7 cm2/s (n=9) and 5.23±1.93x10-7 cm2/s (n=8), respectively. Both diffusion coefficients were insensitive to cell shrinkage caused by ECA during the diffusion experiments and cell damage caused by the storage of tissues ex vivo before the experiments. Binding of ECA to scleral tissues could not be detected. The apparent maximum binding capacity and the apparent equilibrium dissociation constant for NaF were 80±5 mM and 2.5±0.5 mM (n=3), respectively. Conclusions These data demonstrated that ECA diffusion was minimally hindered by structures in the sclera, presumably due to the lack of cells and binding sites for ECA in the sclera. PMID:17356511

Lin, Cheng-Wen; Wang, Yong; Challa, Pratap; Epstein, David L.

2007-01-01

62

Objective area measurement technique for choroidal neovascularization from fluorescein angiography.  

PubMed

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

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

2014-01-01

63

Automated single-slide staining system  

NASA Technical Reports Server (NTRS)

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.

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

1974-01-01

64

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

E-print Network

A series of fluorescein and rosamine derivatives have been prepared and their spectroscopical properties analyzed to determine their usefulness as donor and/or acceptors in "through-bond" energy transfer systems. Such new systems have been tailored...

Castro, Juan C.

2010-07-14

65

INTER-LABORATORY STUDY OF CELLULAR FLUORESCENCE INTENSITY MEASUREMENTS WITH FLUORESCEIN-LABELED MICROBEAD STANDARDS  

EPA Science Inventory

To determine the precision of cellular fluorescence intensity (FI) measurements derived from labeled microbead standards, FI results were compared from 43 different flow cytometers in 34 laboratories. ll laboratories analyzed prepared aliquots of fluoresceinated calf thymocyte nu...

66

Photoactivatable fluorescein derivatives caged with a (3-hydroxy-2-naphthalenyl)methyl group.  

PubMed

The (3-hydroxy-2-naphthalenyl)methyl (NQMP) group represents an efficient photocage for fluorescein-based dyes. Thus, irradiation of the 6-NQMP ether of 2'-hydroxymethylfluorescein with low-intensity UVA light results in a 4-fold increase in emission intensity. Photoactivation of nonfluorescent NQMP-caged 3-allyloxyfluorescein produces a highly emissive fluorescein monoether. To facilitate conjugation of the caged dye to the substrate of interest via click chemistry, the allyloxy appendage was functionalized with an azide moiety. PMID:25036698

Nekongo, Emmanuel E; Popik, Vladimir V

2014-08-15

67

Optos Panoramic200A fluorescein angiography for proliferative diabetic retinopathy with asteroid hyalosis.  

PubMed

We report a unique situation in which the use of Optos Panoramic200A fluorescein angiography directed the management of a patient with asteroid hyalosis and active proliferative diabetic retinopathy. The Optos Panoramic200A system provides a 200 degrees field of view, which may be useful in selected cases where simultaneous view of the posterior pole and periphery is important. The Optos fluorescein angiogram directed the management of our patient with proliferative diabetic retinopathy combined with asteroid hyalosis. PMID:17564923

Win, Peter H; Young, Tara A

2007-01-01

68

Anaphylactic response to topical fluorescein 2% eye drops: a case report  

PubMed Central

Introduction The intravenous use of fluorescein 10% during retinal angiography can cause severe systemic reactions including, on rare occasions, anaphylaxis. Fluorescein 2% eye drops are used extensively for clinical examination and diagnosis, but to the best of our knowledge, they have only been reported as being responsible for a systemic anaphylactic response on two previous occasions. Case presentation We report the case of a 51-year-old woman who developed an anaphylactic reaction when she was administered fluorescein sodium 2% eye drops after cataract surgery. This was the second time she had been exposed to fluorescein. She had brittle asthma and a history of anaphylaxis following exposure to a variety of drug and food allergens. She was successfully resuscitated and recovered completely over a period of two days. Conclusions Fluorescein 2% drops are universally used in general practice, ophthalmology, optometry, and casualty departments. Our case report reveals the potential for this benign eye drop to cause a life-threatening systemic reaction and emphasises the importance of considering this consequence when administering topical fluorescein 2% to a patient with a history of anaphylaxis to other allergens. PMID:20181047

2010-01-01

69

Whole Blood Cell Staining Device  

NASA Technical Reports Server (NTRS)

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.

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

2000-01-01

70

Stain Uses Recipe p-Anisaldehyde General purpose stain,  

E-print Network

Olefins and other readily oxidized groups. Dissolve 1.5 g KMnO4, 10 g K2CO3, and 1.25 mL 10% NaOH in 200 mL water. Cerium Sulfate General stain, particularly useful for alkaloids. Make an aqueous solution of 10% Cerium (IV) sulfate and 15% H2SO4. Morin Hydrate General reagent. Fluorescently active. Make up a 0.1 wt

Yao, Shao Q

71

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

PubMed Central

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

2012-01-01

72

Methods for chromosome-specific staining  

DOEpatents

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.

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

1995-01-01

73

Fluorescein fluorescence use in the management of intracranial neoplastic and vascular lesions: a review and report of a new technique.  

PubMed

The use of fluorescent technologies in neurosurgery has a substantial history with applications to vascular and tumor surgery dating back to the 1940s. This review focuses on the applications of fluorescence imaging to intracranial vascular and neoplastic lesions using sodium fluorescein. The authors performed a literature search for articles about the use of sodium fluorescein in neurosurgery. Fifty-five articles were initially retrieved, and 37 of these were appropriate for this review. The subcategorization of these articles revealed 2 describing the properties of fluorescein, 19 articles relating to applications of fluorescein to tumor, 11 relating to vascular applications, and 5 reporting side effects associated with fluorescein use. Articles related to use of this agent in evaluation of CSF leak were excluded. Sodium fluorescein has been reported to be a useful surgical adjunct in resection of neoplastic lesions based on differential fluorescence between normal and neoplastic tissue. There are many reports on the utility of fluorescein in vascular imaging relating to arteriovenous malformations, aneurysms, and vessel anastomosis; however, these reports do not examine primary outcomes. Sodium fluorescein has been judged as generally safe with few reports of severe complications. Sodium fluorescein has demonstrated promise as a useful surgical adjunct in neurosurgery for vascular and neoplastic lesions. It is well tolerated, but further study is required to determine its full utility. Finally, we will introduce a new practical technology that could potentially improve intraoperative application of sodium fluorescein by improving its fluorescence visualization while using substantially lower doses of this dye. PMID:23363235

Lane, Brandon C; Cohen-Gadol, Aaron A

2013-06-01

74

[Giemsa stain's 100th year].  

PubMed

Research work associated with the development of Giemsa stain is revived, with emphasis on the methylene blue polychromes. A short biographical sketch of Dr. Berthold Gustav Carl Giemsa is presented, along with the composition of his original formulation and its mec. HPLC analyses of his azur II indicated the following composition: methylene blue 63.6%, azur B 28.6%, azur A 4.4%, azur C 1.4%, thionin 1.9%. Azur I was not "pure", but rather a mixture of thionin and all of its 3 and 7 N-methylated derivatives. Lillie inferred that it was probably prepared by an acid oxidation process. Applications of Giemsa stain reported in the last 32 years are tabulated. PMID:12696395

Perea-Sasiaín, José

2003-03-01

75

Double-staining epifluorescence technique to assess frequency of dividing cells and bacteriovory in natural populations of heterotrophic microprotozoa.  

PubMed

We have developed a double-staining procedure for use with epifluorescence microscopy which allows the detection both of dividing cells and of ingested bacteria in food vacuoles of heterotrophic microprotozoa. Microprotozoan cells are stained sequentially with the DNA-specific fluorochrome DAPI (4',6-diami-dino-2-phenylindole) and the nonspecific protein stain fluorescein isothiocyanate. During microscopic examination, heterotrophic microprotozoan cells are first located with fluorescein isothiocyanate fluorescence and then epifluorescence filter sets are switched to permit inspection under DAPI fluorescence of the cell nuclei and of the contents of food vacuoles. Among in situ populations of estuarine microprotozoa sampled over a tidal cycle, we found from 2.2 to 5.2% of the heterotrophic cells in a recognizable stage of division (nuclei elongated or double). Batch culture growth experiments were also carried out both with natural populations and with two isolated species of estuarine microprotozoa. In these experiments, the frequency of dividing cells ranged from 1.2 to 3.8% and appeared to be negatively correlated with growth rate. Microprotozoan populations sampled in continental shelf waters off Savannah, Ga., had mean frequencies of dividing cells ranging from 2.0 to 5.0%. A large fraction of cells in heterotrophic microprotozoan populations (an average of 27.4 +/- 1.0% in estuarine water and of 30.1 +/- 4.8% in shelf water) had DAPI-stained inclusions, presumably recently ingested bacteria, in their food vacuoles. PMID:16346446

Sherr, E B; Sherr, B F

1983-12-01

76

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

PubMed

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

Noga, E J; Udomkusonsri, P

2002-11-01

77

One-Pot Catalytic Conversion of Epoxides to 1,2-Diacetates with Hydride Transferring Agents in Acetic Anhydride  

Microsoft Academic Search

Direct transformation of structurally different epoxides to the corresponding 1,2-diacetates was studied with catalytic amounts of NaBH4, LiAlH4, CaH2, and NaH. The reactions were carried out in refluxing acetic anhydride within 1.5–2.5 h to give vic-diacetates in good to excellent yields. Conversion of R-(+)-styrene oxide to S-(+)-1,2-diacetoxy-1-phenylethane was carried out with good yield and stereospecificity with the NaBH4\\/Ac2O system at 0 °C.

Behzad Zeynizadeh; Leila Sadighnia

2011-01-01

78

Problem Solving: Pencil Box Staining  

NSDL National Science Digital Library

This professional development video clip shows students engaged in the first Common Core Practice StandardâMake sense of problems and persevere in solving them as learners make a decision about how much stain will be needed to cover the surface area of twenty-six completed boxes. Additional resources include a video transcript, teaching tips, and a link to a professional development reflection activity based upon the video. A related clip (cataloged separately) shows the same exploration by the same students but Common Core Practice Standard # #5-Use appropriate tools strategically is evident.

Boston, Wghb

2013-01-01

79

Safer staining method for acid fast bacilli.  

PubMed Central

To develop a method for staining acid fast bacilli which excluded highly toxic phenol from the staining solution. A lipophilic agent, a liquid organic detergent, LOC High Studs, distributed by Amway, was substituted. The acid fast bacilli stained red; nuclei, cytoplasm, and cytoplasmic elements stained blue on a clear background. These results compare very favourably with acid fast bacilli stained by the traditional method. Detergents are efficient lipophilic agents and safer to handle than phenol. The method described here stains acid fast bacilli as efficiently as traditional carbol fuchsin methods. LOC High Suds is considerably cheaper than phenol. Images PMID:7687254

Ellis, R C; Zabrowarny, L A

1993-01-01

80

Safer staining method for acid fast bacilli.  

PubMed

To develop a method for staining acid fast bacilli which excluded highly toxic phenol from the staining solution. A lipophilic agent, a liquid organic detergent, LOC High Studs, distributed by Amway, was substituted. The acid fast bacilli stained red; nuclei, cytoplasm, and cytoplasmic elements stained blue on a clear background. These results compare very favourably with acid fast bacilli stained by the traditional method. Detergents are efficient lipophilic agents and safer to handle than phenol. The method described here stains acid fast bacilli as efficiently as traditional carbol fuchsin methods. LOC High Suds is considerably cheaper than phenol. PMID:7687254

Ellis, R C; Zabrowarny, L A

1993-06-01

81

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

PubMed Central

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

Flemmig, Jorg; Zschaler, Josefin; Remmler, Johannes; Arnhold, Jurgen

2012-01-01

82

Structural studies of the mechanism for biosensing antibiotics in a fluorescein-labeled ?-lactamase  

PubMed Central

Background ?-lactamase conjugated with environment-sensitive fluorescein molecule to residue 166 on the ?-loop near its catalytic site is a highly effective biosensor for ?-lactam antibiotics. Yet the molecular mechanism of such fluorescence-based biosensing is not well understood. Results Here we report the crystal structure of a Class A ?-lactamase PenP from Bacillus licheniformis 749/C with fluorescein conjugated at residue 166 after E166C mutation, both in apo form (PenP-E166Cf) and in covalent complex form with cefotaxime (PenP-E166Cf-cefotaxime), to illustrate its biosensing mechanism. In the apo structure the fluorescein molecule partially occupies the antibiotic binding site and is highly dynamic. In the PenP-E166Cf-cefatoxime complex structure the binding and subsequent acylation of cefotaxime to PenP displaces fluorescein from its original location to avoid steric clash. Such displacement causes the well-folded ?-loop to become fully flexible and the conjugated fluorescein molecule to relocate to a more solvent exposed environment, hence enhancing its fluorescence emission. Furthermore, the fully flexible ?-loop enables the narrow-spectrum PenP enzyme to bind cefotaxime in a mode that resembles the extended-spectrum ?-lactamase. Conclusions Our structural studies indicate the biosensing mechanism of a fluorescein-labelled ?-lactamase. Such findings confirm our previous proposal based on molecular modelling and provide useful information for the rational design of ?-lactamase-based biosensor to detect the wide spectrum of ?-lactam antibiotics. The observation of increased ?-loop flexibility upon conjugation of fluorophore may have the potential to serve as a screening tool for novel ?-lactamase inhibitors that target the ?-loop and not the active site. PMID:21443768

2011-01-01

83

Fluorogenic Substrate Detection of Viable Intracellular and Extracellular Pathogenic Protozoa  

NASA Astrophysics Data System (ADS)

Viable Leishmania promastigotes and amastigotes were detected by epifluorescence microscopy with fluorescein diacetate being used to mark living parasites and the nucleic acid-binding compound ethidium bromide to stain dead cells. This procedure is superior to other assays because it is faster and detects viable intracellular as well as extracellular Leishmania. Furthermore, destruction of intracellular pathogens by macrophages is more accurately determined with fluorescein diacetate than with other stains. The procedure may have applications in programs to develop drugs and vaccines against protozoa responsible for human and animal disease.

Jackson, Peter R.; Pappas, Michael G.; Hansen, Brian D.

1985-01-01

84

Methods for chromosome-specific staining  

DOEpatents

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.

Gray, J.W.; Pinkel, D.

1995-09-05

85

[Progress in the treatment of stained teeth].  

PubMed

The treatment of stained teeth has been one of the striking aspects of stomatology and esthetic dentistry. Based on detailed data and references, this article introduces the types of stained teeth and the main treatment methods including strong point, weakness, limitation of the usage, result, and the relevant mechanisms. It addresses the researches on problems in the treatment of stained teeth. Also in this paper is envisaged what will be done to treat the stained teeth in future. PMID:15250169

Huo, Danqun; Xie, Guo; Hou, Changjun; Liu, Jia; Huang, Chunyue; He, Zuoyun

2004-06-01

86

An Accelerated Method for Staining Tularemia Bacteria.  

National Technical Information Service (NTIS)

At present, the standard procedure for staining tularemia bacteria in animal tissue is the staining of the smear-impression according to Romanovskiy-Gimze. In a search for a more optimal method, the author decided on a method of accelerating staining whic...

R. I. Kudelina

1978-01-01

87

Original article Blue-stain fungi associated  

E-print Network

Original article Blue-stain fungi associated with Tomicus piniperda in Sweden and preliminary to determine the development of blue-staining of sapwood. Fungi were isolated from samples of inner bark and blue-stained sapwood in connection with galleries of T piniperda. Samples were also taken from beetle

Paris-Sud XI, Université de

88

Cellulose diacetate-g-poly(p-dioxanone) co-polymer: synthesis, properties and microsphere preparation.  

PubMed

A novel biodegradable graft co-polymer based on cellulose diacetate (CDA) and poly(p-dioxanone) was synthesized by ring-opening polymerization of p-dioxanone (PDO). The molecular structure of co-polymers was characterized by one- and two-dimensional NMR. The graft co-polymers with different lengths of PPDO side-chains could be controllably synthesized by changing the in-feed ratio of CDA/PDO. It was found that the PPDO content had great effect on the thermal transition behavior, crystallization ability and thermal stability of the graft co-polymer. The in vitro degradation rates of CDA-g-PPDO were higher than that of linear PPDO due to their lower crystallinity. Moreover, porous microspheres of graft co-polymers with a diameter of about 5 ?m, prepared through the solvent evaporation of water-in-oil emulsion (W/O), indicated it may have potential applications in drug-delivery systems. PMID:20566069

Zhu, Jiang; Dang, Hai-Chun; Wang, Wen-Tao; Wang, Xiu-Li; Wang, Yu-Zhong

2011-01-01

89

Microdissection of stained archival tissue.  

PubMed Central

In many tissues the preinvasive stage of neoplastic progression can be identified histologically as dysplasia or in situ disease. There is much interest in defining the molecular events associated with the early stages of neoplasia. Retrieval of histologically recognisable preinvasive neoplastic tissue uncontaminated by inflammatory or stromal cells is important for genetic studies using polymerase chain reaction (PCR) assay. A novel method for microdissection is described in which 10 microns sections are dewaxed, stained with haematoxylin and eosin, dried, covered with Sellotape, and the tissue cut out using a scalpel blade under direct visual control. The method is quick, eliminates problems of operator tremor, preserves the architecture of the micro-dissected tissue (for photographic documentation) and requires no special equipment. The presence of Sellotape and adhesive in the reaction mixture has no detrimental effect on the ability to extract DNA or to perform PCR. Images PMID:9350307

Gupta, S K; Douglas-Jones, A G; Morgan, J M

1997-01-01

90

Correlation of electroretinographic and fluorescein angiographic findings in unilateral central retinal vein obstruction  

Microsoft Academic Search

• Background: In central retinal vein obstruction (CRVO), electroretinogram (ERG) abnormalities and extensive retinal capillary dropout (CD) in the fluorescein angiogram (FA) are good indicators of retinal ischemia. We retrospectively studied patients with unilateral CRVO and compared the ERG and FA results • Methods: Single white flash ERG, photopic ERG, scotopic ERG and flicker ERG were recordered in 30 cases

Yoshie Matsui; Osamu Katsumi; Mehul C. Mehta; Tatsuo Hirose

1994-01-01

91

Fluorescein angiography and its prognostic significance in central retinal vein occlusion  

Microsoft Academic Search

To determine the prognostic value of fluorescein angiograms in central retinal occlusion 75 patients presenting within the first three months of their initial visual symptoms were studied prospectively. The prognosis was found to be best (complete or partial resolution) in eyes with good capillary perfusion, an intact perifoveal capillary arcade, and leakage only from terminal venules and veins. A broken

L Laatikainen; E M Kohner

1976-01-01

92

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

NASA Astrophysics Data System (ADS)

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

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

2013-10-01

93

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

PubMed Central

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

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

94

Ultraphosphate, a potent stain control agent that is effective for both stain removal and prevention of stain deposition.  

PubMed

Polyphosphate is a phosphate polymer which is effective for stain removal and prevention of stain deposition. Ultraphosphate belongs to the polyphosphate group and has a highly branched mesh-like structure. To evaluate stain control ability of ultraphosphate, we used HAP powder, glass-ionomer cement and detached human teeth for models of in vitro stain control experiments. When using HAP powder, the stain removal ability of ultraphosphate was the highest among common chelating agents. In addition, ultraphosphate efficiently removed stain and prevented stain deposition on glass-ionomer cement at 20°C and 37°C. Finally, ultraphosphate removed coffee stain from human teeth surface efficiently and the color difference (?E*ab) before and after ultraphosphate treatment was changed dramatically from 59.4 to 8.3. Similarly, the ?E*ab value of human teeth treated with ultraphosphate before coffee treatment was only 9.9, while the value without ultraphosphate pre-treatment was 21.2. These results indicate that ultraphosphate is a potent agent for stain control. PMID:24598236

Koyasu, Masahiro; Shiba, Toshikazu; Kawazoe, Yumi; Manabe, Atsufumi; Miyazaki, Takashi

2014-01-01

95

Staining sectioned biological specimens for transmission electron microscopy: conventional and en bloc stains.  

PubMed

Post-staining of ultrathin sections and/or en bloc staining of specimens is necessary for differential contrast and improved resolution of cellular structures. Often specimens are fixed and stained with osmium tetroxide during fixation, but additional contrast is the result of additional heavy metal stains on the sections. The most common post-staining of sections is done on grids by aqueous uranyl acetate followed by lead citrate. When it is apparent that simple, aqueous uranium and lead post-staining is not adequate, other stains are invoked. These procedures can be as simple as en bloc staining with uranyl acetate after primary fixation and osmication. Over the years, several other treatments have been developed for use with the primary fixation or during dehydration. Tannic acid, paraphenylenediamine (PPD), and malachite green can all serve as en bloc stains and can contribute to overall improved visualization of ultrastructural details in biological specimens. Tannic acid and PPD improve membrane preservation, and malachite green is a phospholipid stain. All of these stains are compatible with aqueous fixatives and should be considered when the usual stains are not satisfactory. Marinozzi rings and microwave-assisted post-staining offer alternatives to traditional grid staining. In addition, stain precipitates on grids often can be removed by treatment with 10 % (v/v) acetic acid. PMID:24357359

Ellis, E Ann

2014-01-01

96

Immune response to a hapten of fluorescein isothiocyanate in a single mouse analyzed by two-dimensional affinity electrophoresis.  

PubMed

Immune response to a hapten of fluorescein isothiocyanate (FITC) in a single BALB/c strain mouse was analyzed by two-dimensional affinity electrophoresis (2D-AEP). Anti-FITC antibodies were induced by immunization with FITC-conjugated bovine serum albumin. The antibodies were separated into a large number of spots of IgG due to differences in their isoelectric points(pI) and binding affinities to the FITC ligand. These spots consisted of IgG families which were composed of several spots having an identical affinity to the ligand but a different pI. The spots were not clearly detected in the antiserum taken on day 7 after the primary immunization, but on day 21 the spots of IgG were clearly detected, with a high diversity and specificity for the ligand. The size and number of IgG spots were markedly increased by the secondary immunization; however, the third immunization did not increase the size and number of IgG spots. The IgG spots of each family were specifically stained with an antimouse IgG subclass antibody. Furthermore, a monoclonal antibody (FL-D6) was separated by 2D-AEP into a single family which consisted of seven IgG1 spots having an identical affinity to FITC but different pIs. Therefore, each of the IgG families of anti-FITC antibodies in the antiserum can be generated by a single clone of anti-FITC antibody-producing cells. The substitution of dextran T2000 or lipopolysaccharide for bovine serum albumin as a carrier for FITC induced much smaller amounts of anti-FITC antibodies with a low diversity but high specificity to FITC. PMID:8462521

Nakamura, K; Mimura, Y; Takeo, K

1993-01-01

97

Stain Removal from a Silicone Maxillofacial Elastomer  

Microsoft Academic Search

In this study, environmental stains were removed from maxillofacial elastomers by solvent extraction. Silastic 44210, an RTV silicone with proven color and physical property stability, was stained with lipstick, disclosing solution, and methylene blue. These stains were then removed by solvent extraction with each of four chemically dissimilar solvents, namely: toluene, benzene, 1,1,1-trichloroethane, and n-hexane. An additional series of samples

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

1981-01-01

98

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

PubMed

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

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

2014-10-01

99

Detection of hyaluronidase activity using fluorescein labeled hyaluronic acid and fluorescence correlation spectroscopy  

PubMed Central

The over-expression of hyaluronidase has been observed in many types of cancer, suggesting that it may have utility for diagnosis. Here we present a technique for the detection of hyaluronidase using Fluorescence Correlation Spectroscopy (FCS). Hyaluronan macromolecules (HAs) have been heavily labeled with fluorescein amine resulting in strong self-quenching. In the presence of hyaluronidase, HA is cleaved into smaller, fluorescein-labeled fragments and the self-quenching is released. Such cleavage is manifested by the increased average diffusion rate of the HA fragments, increased concentration of individual, fluorescent HA fragments, and increased intensity. All three of these properties are monitored simultaneously throughout FCS measurements, both as a function of time and hyaluronidase concentration. The method we present provides a sensitive measure of hyaluronidase activity and requires extremely small amounts of the HA substrate. PMID:23018154

Rich, Ryan M.; Mummert, Mark; Foldes-Papp, Zeno; Gryczynski, Zygmunt; Borejdo, Julian; Gryczynski, Ignacy; Fudala, Rafal

2012-01-01

100

Effect of silver NPs plasmon on optical properties of fluorescein dye  

NASA Astrophysics Data System (ADS)

In this work we studied the effect of silver nanoparticles "AgNPs" on the optical properties of fluoretain-->rescein dye. Fluorescein dye solutions have been mixed with different concentrations of colloidal AgNPs. Absorption and fluorescence enhancement of fluorescein dye molecules was detected in the presence of AgNPs. Fluorescence enhancement of the dye molecules was observed with a maximal enhancement factor of about 3-fold. Enhancement of the rate of radiative transition was also detected. The enhancement mechanisms are attributed to a modification of the local density of electromagnetic modes in the vicinity of AgNPs at energies resonant with surface Plasmon. The ability of fluorophore-metal mixture to actively enhance the dye's luminescence could leads to new opportunities for technological development of light emitting and photonic devices. It also may have applications in the fields of bio-technology and medical diagnostics as new class of fluorescence based sensing.

Ragab, Alaa EL-din E. A.; Gadallah, A.; Mohamed, Mona B.; Azzouz, I. M.

2013-11-01

101

Influence of ionization states of antigen on anti-fluorescein antibodies  

NASA Astrophysics Data System (ADS)

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.

Fukunishi, Hiroaki

2012-10-01

102

Investigation of the hydrogen-bond structure of cellulose diacetate by two-dimensional infrared correlation spectroscopy  

Microsoft Academic Search

Temperature-dependent structural changes in hydrogen bonds in cellulose diacetate (CDA) were investigated by Fourier transform infrared spectroscopy (FT-IR). The OH stretching vibration band was selected to explore the structure changes. Two-dimensional correlation spectroscopy (2DCOS) in combination with moving-window technique was applied to analyze the overlapping OH band due to various kinds of hydrogen bonds. By virtue of this powerful method,

Yilu Guo; Peiyi Wu

2008-01-01

103

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

PubMed Central

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

Piotrowiak, Ilona; Kaluzny, Bartlomiej J.; Danek, Beata; Chwiedacz, Adam; Sikorski, Bartosz L.; Malukiewicz, Grazyna

2014-01-01

104

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

NASA Astrophysics Data System (ADS)

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.

Zhang, Li; Zhang, Xianhong

2014-12-01

105

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

PubMed

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

Zhang, Li; Zhang, Xianhong

2014-12-10

106

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

NASA Astrophysics Data System (ADS)

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.

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

2014-12-01

107

Divided we stand: tracking cell proliferation with carboxyfluorescein diacetate succinimidyl ester.  

PubMed

Most techniques for assessing cell division can either detect limited numbers of cell divisions (bromodeoxyuridine incorporation) or only quantify overall proliferation (tritiated thymidine incorporation). In the majority of cases, viable cells of known division history cannot subsequently be obtained for functional studies. The cells of the immune system undergo marked proliferation and differentiation during the course of an immune response. The relative lack of an organized structure of the lymphohaemopoietic system, in contrast with other organ systems, makes lineage interrelationships difficult to study. Coupled with the remarkable degree of mobility engendered by recirculation, the differentiation occurring along with cell division in the immune system has not been readily accessible for investigation. The present article reviews the development of a cell division analysis procedure based on the quantitative serial halving of the membrane permeant, stably incorporating fluorescent dye carboxyfluorescein diacetate succinimidyl ester (CFSE or CFDA, SE). The technique can be used both in vitro and in vivo, allowing eight to 10 successive divisions to be resolved by flow cytometry. Furthermore, viable cells from defined generation numbers can be sorted by flow cytometry for functional analysis. PMID:10571671

Lyons, A B

1999-12-01

108

Standardization of the Papanicolaou stain. I. A comparison of five nuclear stains.  

PubMed

The staining characteristics of five nuclear stains used in a Papanicolaou staining procedure were investigated. Alcohol-fixed cervical smears were stained with a modified Papanicolaou procedure using hematoxylin, alcoholic thionin bromide, alcoholic Victoria blue B, gallocyanin or the thionin Feulgen reagent (thionin-SO2) as the nuclear stain. The same anionic counterstain was used for all slides, and the optical densities of cell nuclei and cytoplasm were measured with the IBAS 2000 image analyzer. Alcoholic thionin gave the most intense nuclear stain, with a very high reproducibility of the staining pattern. Hematoxylin showed the highest coefficient of variation of the staining intensity. Both hematoxylin and gallocyanin gave some nonspecific cytoplasmic staining. Thionin-SO2 allowed a quantitative assessment of DNA, but gave a low staining intensity. Staining with the metal complex dyes interfered with subsequent staining with the pararosaniline Feulgen reagent. Alcoholic thioinin is thus recommended as a nuclear stain for cervical cytology in the Papanicolaou procedure, both for image analysis and for visual microscopy. PMID:1695100

Schulte, E; Wittekind, D

1990-06-01

109

Purified azure B as a reticulocyte stain  

Microsoft Academic Search

A comparison has been made between reticulocyte preparations stained with purified azure B and with several commerically available batches of brilliant cresyl blue and new methylene blue. Marked variations were observed in the composition and staining performances of the various batches of the two commerically available dyes. Although there were no significant differences in reticulocyte counts obtained with these two

P N Marshall; S A Bentley; S M Lewis

1976-01-01

110

Fluorescein angiography  

MedlinePLUS

... bar to keep your head still during the test. The health care provider will take pictures of the inside of ... form. You must remove contact lenses before the test. Tell the health care provider if you may be pregnant.

111

Negative staining and Cryo-negative Staining of Macromolecules and Viruses for TEM  

PubMed Central

In this review we cover the technical background to negative staining of biomolecules and viruses, and then expand upon the different possibilities and limitations. Topics range from conventional air-dry negative staining of samples adsorbed to carbon support films, the variant termed the “negative staining-carbon film” technique and negative staining of samples spread across the holes of holey carbon support films, to a consideration of dynamic/time-dependent negative staining. For each of these approaches examples of attainable data are given. The cryo-negative staining technique for the specimen preparation of frozen-hydrated/vitrified samples is also presented. A detailed protocol to successfully achieve cryo-negative staining with ammonium molybdate is given, as well as examples of data, which support the claim that cryo-negative staining provides a useful approach for the high-resolution study of macromolecular and viral structure. PMID:20634082

De Carlo, Sacha; Harris, J. Robin

2010-01-01

112

Compact, Automated Centrifugal Slide-Staining System  

NASA Technical Reports Server (NTRS)

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

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

2004-01-01

113

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

PubMed

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

Smith, A A; Glickfield, I

2011-04-01

114

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

EPA Science Inventory

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

115

COMPARISON OF ANIMAL INFECTIVITY, EXCYSTATION AND FLUOROGENIC DYE AS MEASURES OF GIARDIA MURIS CYST INACTIVATION BY OZONE  

EPA Science Inventory

Giardia muris cyst viability following ozonation was compared using fluorescein diacetate-ethidium bromide staining, the C3H/HeN mouse-G. muris model, and in vitro excystation. ench-scale, batch experiments were conducted under laboratory conditions (pH 6.7; 22 degrees C) in ozon...

116

Gram staining apparatus for space station applications  

NASA Technical Reports Server (NTRS)

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.

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

1990-01-01

117

Gram staining apparatus for space station applications.  

PubMed

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. PMID:1690529

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

1990-03-01

118

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

PubMed Central

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

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

119

Characterization of surface sugars on algal cells with fluorescein isothiocyanate-conjugated lectins  

Microsoft Academic Search

Summary.  We used qualitative and quantitative fluorescence microscopy of the fluorescein isothiocyanate-conjugated lectins Concanavalin\\u000a A, phytohaemagglutinin-erythroagglutinin, pokeweed mitogen, and peanut agglutinin to examine sugar composition on the cell\\u000a surface and cell-associated mucilage (where present) in a number of cultured and environmental algae. Lectin-binding activity\\u000a was markedly different between laboratory-cultured and environmental samples of the same species. Sugar composition of the\\u000a cyanobacterium

C.-J. Tien; D. C. Sigee; K. N. White

2005-01-01

120

Indium–tin-oxide thin films prepared by dip-coating of indium diacetate monohydroxide and tin dichloride  

Microsoft Academic Search

Tin-doped In2O3 (ITO) films were prepared by the dip-coating method using an ethanol solution of indium diacetate monohydroxide, In(OH)(CH3COO)2, and tin dichloride, SnCl2·2H2O, with 2-aminoethanol (monoethanolamine), H2NC2H4OH. The influence of the tin concentration was investigated between 0 and 20 at.% Sn. The composition of the films (thickness ?90 nm) approximately agreed with that of the solution when the dipping and

Shigeyuki Seki; Yutaka Sawada; Toshikazu Nishide

2001-01-01

121

Stain Removal from a Pigmented Silicone Maxillofacial Elastomer  

Microsoft Academic Search

The removal of environmental stains from a pigmented maxillofacial elastomer was carried out by solvent extraction under network swelling. Silastic 44210 was pigmented with 11 maxillofacial pigments prior to staining. Samples were stained with lipstick, methylene blue, and disclosing solution. These stains were then removed by solvent extraction with 1,1,1-trichloroethane. Color parameter measurements both before and after staining and after

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

1982-01-01

122

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

PubMed Central

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

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

2013-01-01

123

Compositions for chromosome-specific staining  

DOEpatents

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.

Gray, J.W.; Pinkel, D.

1998-05-26

124

Compositions for chromosome-specific staining  

DOEpatents

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.

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

1998-01-01

125

Resistance to Extrinsic Stains by Hydrophobic Composite Resin Systems  

Microsoft Academic Search

Measurement of color parameters demonstrated that hydrophobic composites had less staining capacity and greater ease of stain removal when compared with conventional composite materials. Accelerated aging of samples is important in staining tests.

W. H. Douglas; R. G. Craig

1982-01-01

126

Automated single-slide staining device  

NASA Technical Reports Server (NTRS)

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.

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

1977-01-01

127

Detection Of Concrete Deterioration By Staining  

DOEpatents

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.

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

1999-09-21

128

Continuous-flow cytomorphological staining and analysis.  

PubMed

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

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

2014-02-01

129

The Language of Stained-Glass Windows  

ERIC Educational Resources Information Center

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…

Brew, Charl Anne

2010-01-01

130

Method for copper staining of germanium crystals  

NASA Technical Reports Server (NTRS)

Proper conditions for copper staining of germanium crystals include a low solution temperature of 3 degrees C, illumination of the sample by infrared light, and careful positioning of the light source relative to the sample so as to minimize absorption of the infrared light.

Rivet, E. J.

1969-01-01

131

Solid acid catalysts: Stain and shine  

NASA Astrophysics Data System (ADS)

Catalyst particles for fluid catalytic cracking are vital for the oil-refinery industry, but their activity is hard to diagnose because of their inter- and intra-particle structural inhomogeneity. With fluorescence confocal microscopy and selective staining, one can now pinpoint the catalytic activity within single catalyst particles from an industrial reactor.

Chen, Peng

2011-11-01

132

Quantitative spatiotemporal image analysis of fluorescein angiography in age-related macular degeneration  

NASA Astrophysics Data System (ADS)

Interpretation and analysis of retinal angiographic studies has been largely qualitative. Quantitative analysis of pathologic fundus features will facilitate interpretation and potentiate clinical studies where precise image metrology is vital. Fluorescein angiography studies of patients with age- related macular degeneration were digitized. Sequential temporal images were spatially-registered with polynomial warping algorithms, allowing for the construction of a three- dimensional (two spatial and one temporal) angiogram vector. Temporal profiles through spatially-registered, temporally- sequential pixels were computed. Characteristic temporal profiles for fundus background, retinal vasculature, retinal pigment epithelial atrophy, and choroidal neovascular (CNV) membranes were observed, allowing for pixel assignment and fundus feature quantitation. Segmentation and quantitation of fundus features including geographic atrophy and CNV is facilitated by spatio-temporal image analysis.

Berger, Jeffrey W.

1998-06-01

133

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

SciTech Connect

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.

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

1981-02-01

134

The fluorescein disappearance test (FDT): an evaluation of its use in infants.  

PubMed

We report a prospective evaluation of the fluorescein disappearance test (FDT) in the diagnosis of lacrimal outflow obstruction in infants under 1 year of age. In a preliminary study of 80 lacrimal systems, we showed that the FDT is a sensitive (90%) and specific (100%) test in aiding the diagnosis of nasolacrimal obstruction in infants. In the main study, we examined 288 eyes of 232 children with epiphora. The FDT was abnormal in 237 (82%). Of the remaining 51 eyes, there was a clinical explanation for this in 33 cases (epiblepharon, conjunctivitis, etc). We have found that this simple and reliable test taken in conjunction with clinical examination provides an objective assessment of the lacrimal outflow status of young children. PMID:1757852

MacEwen, C J; Young, J D

1991-01-01

135

Hydroquinone-O,O?-diacetic acid as a more labile replacement for succinic acid linkers in solid-phase oligonucleotide synthesis  

Microsoft Academic Search

Hydroquinone-O,O?-diacetic acid (QDA) can be used to link nucleosides to CPG or polystyrene supports instead of succinic acid. Cleavage of oligodeoxy- or oligoribonucleotides, using ammonium hydroxide, requires only two to five minutes. The QDA linker is stable, easily prepared and does not require any other changes to the reagents, methods, or instrumentation used in automated solid-phase oligonucleotide synthesis.

Richard T. Pon; Shuyuan Yu

1997-01-01

136

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

E-print Network

68Ga-N,N'-bis[2-Hydroxy-5-(carboxyethyl)benzyl] ethylenediamine-N,N'-diacetic acid-polyethylene glycol-single- chain Cys-tagged vascular endothelial growth factor-121 [68Ga-HBED-CC-PEG-scVEGF] Kam Leung, PhD National Center for Biotechnology Information, NLM, NIH, Bethesda, MD Chemical name: 68Ga

Levin, Judith G.

137

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

NASA Astrophysics Data System (ADS)

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

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

2014-03-01

138

Silver Staining of 2D Electrophoresis Gels Thierry Rabilloud  

E-print Network

Silver Staining of 2D Electrophoresis Gels Thierry Rabilloud CEA-DSV-iRTSV/LCBM and UMR CNRS Head: Silver staining #12;i. Abstract Silver staining is used to detect proteins after electrophoretic are discussed in this chapter, and optimized silver staining protocols are proposed. ii. Key Words: mass

Paris-Sud XI, Université de

139

Inhibition of nonproteolytic, psychrotrophic clostridia and anaerobic sporeformers by sodium diacetate and sodium lactate in cook-in-bag turkey breast.  

PubMed

A nonproteolytic, psychrotrophic Clostridium isolate, designated strain OMFRI1, was recovered from cook-in-bag turkey breasts (CIBTB) that displayed an intense pink discoloration and an off-odor following extended refrigerated storage. The viability of strain OMFRI1 in CIBTB containing sodium diacetate (at 0, 0.25, and 0.5%) and/or sodium lactate (at 0, 1.25, and 2.5%) was subsequently evaluated. Raw CIBTB batter was inoculated with 9 to 30 spores of strain OMFRI1 per g, vacuum packaged, cooked to an instantaneous internal temperature of 71.1 degrees C, chilled, and incubated at 4 degrees C for up to 22 weeks. In the absence of food-grade antimicrobial agents, spoilage (i.e., an off-odor) occurred within 6 weeks, and anaerobic plate counts reached 6.6 log10 CFU/g. The CIBTB containing sodium diacetate (0.25%) and that containing sodium lactate (1.25%) required 12 weeks for spoilage to occur and for anaerobic plate counts to reach 7.0 and 6.0 log10 CFU/g, respectively. When sodium diacetate (0.25%) and sodium lactate (1.25%) were used in combination, no off-odor was detected and anaerobic plate counts did not exceed 2.3 log10 CFU/g over 22 weeks of storage at 4 degrees C. In related experiments, sodium diacetate (at 0, 0.25, and 0.5%), sodium lactate (at 0, 1.25, and 2.5%), and combinations of both ingredients were evaluated in uninoculated CIBTB incubated at 25 degrees C for up to 22 days. In the absence of antimicrobial agents and in CIBTB containing sodium diacetate (0.5%), spoilage occurred within 8 days and anaerobic plate counts reached 6.8 and 6.6 log10 CFU/g, respectively. Samples of CIBTB containing sodium lactate (2.5%) showed signs of spoilage within 22 days, and anaerobic plate counts for these samples ranged from < or = 1.0 to 6.3 log10 CFU/g. In CIBTB containing both sodium lactate (2.5%) and sodium diacetate (0.25%), spoilage was not evident and anaerobic plate counts were < or = 1.0 log10 CFU/g within 22 days. These data validate the efficacy of sodium lactate and sodium diacetate in extending the shelf life of CIBTB. PMID:12929840

Meyer, J D; Cerveny, J G; Luchansky, J B

2003-08-01

140

Comparison of SF6 and Fluorescein as Tracers for Measuring Transport Processes in a Large Tidal River  

E-print Network

conducted in the tidal Hudson River. At the beginning of the experiment, 36 kg of fluorescein and 4.3 mol of SF6 were injected into the Hudson River at an averaged depth of 9.5 m, 1 m above the bottom, near: Advection; Dispersion; Mixing; Tracers; Dyes; Hudson River; Tides. Introduction Net advection

Ho, David

141

Newer applications of the histological stain prepared from Pterocarpus santalinus.  

PubMed

A histological stain prepared from the heartwood of Pterocarpus santalinus Linn. has been found to be an excellent nuclear stain for various cells of animal and plant origin. As an elastic tissue stain, the results are comparable to standard elastic tissue stains. The striations of voluntary muscle fibers are well shown. The Nissl granules and fibers of cranial nerves in the pons are visualized. When counterstained with light green, it differentially stains muscle and fibrous tissue. The stain can be used as counterstain with certain histochemical procedures with satisfactory results. The preparation and use of this versatile stain are described. PMID:6166099

Sen Gupta, P C; Mukherjee, A K

1981-03-01

142

[Modification of Harada's method for rapid staining of mycobacteria].  

PubMed

Harada employed periodic acid-carbol pararosaniline and periodic acid-methenamine silver stain for demonstrating chromophobic bacilli which do not get stained with conventional carbol fuchsin or counter stain. This staining method takes considerable time for complete oxidation with periodic acid. We have succeeded in reducing the oxidation time by using hydrogen peroxide treatment prior to periodic acid and with the use of acidified sodium hydrogen sulfite treatment before carbol pararosaniline stain. We also found that in methenamine silver stain, combined use of semicarbazide and microwave treatment can shorten the whole staining time up to four hours without losing ito sensitivity. PMID:1284986

Kawatsu, K; Izumi, S; Yumi, M; Butt, K I; Wang, T

1992-11-01

143

FISH and immunofluorescence staining in Chlamydomonas.  

PubMed

Here we describe how to use fluorescence in situ hybridization and immunofluorescence staining to determine the in situ distributions of specific mRNAs and proteins in Chlamydomonas reinhardtii. This unicellular eukaryotic green alga is a major model organism in cell biological research. Chlamydomonas is well suited for these approaches because one can determine the cytological location of fluorescence signals within a characteristic cellular anatomy relative to prominent cytological markers. Moreover, FISH and IF staining offer practical alternatives to techniques involving fluorescent proteins, which are difficult to express and detect in Chlamydomonas. The main goal of this review is to describe these powerful tools and to facilitate their routine use in Chlamydomonas research. PMID:21431732

Uniacke, James; Colón-Ramos, Daniel; Zerges, William

2011-01-01

144

The spots and stains of plate tectonics  

NASA Astrophysics Data System (ADS)

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.

Oliver, Jack

1992-01-01

145

Laser Treatment of Port Wine Stains  

NASA Astrophysics Data System (ADS)

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.

Majaron, Boris; Nelson, J. Stuart

146

Hematoxyliin & Eosin Staining 1. Xylene 3 min  

E-print Network

Hematoxyliin & Eosin Staining 1. Xylene 3 min 2. Xylene up & down 3. 100% EtOH up & down 4. 100% Et min, until no more schlieren 9. Distilled H2O dip 10. Hematoxylin 2a 3 min 11. Running tap water 2 min 12. Acid EtOHb 2 quick dips then slowly to 14 13. Running tap water 1-2 min, repeat above if still

Oliver, Douglas L.

147

Activity staining of endoglucanases in polyacrylamide gels.  

PubMed

The endoglucanases of Penicillium funiculosum were analyzed for the presence of multiple forms using a modified version of the Congo red method. Postelectrophoretic slab gels were directly incubated in a solution of carboxymethylcellulose for a period as short as 15 min and then the activities were visualized by staining with Congo red. Ten distinct bands of clearances were obtained indicating the presence of at least as many multiple forms. PMID:1280921

Mathew, R; Rao, K K

1992-10-01

148

Hydroxychloroquine-induced hyperpigmentation: the staining pattern.  

PubMed

We report two cases of hydroxychloroquine-induced hyperpigmentation presenting in a 50-year-old Caucasian female (case 1) and a 78-year-old female (case 2), both receiving 400 mg per day. Case 1 had an arthritis predominant undifferentiated connective tissue disease, which was treated with hydroxychloroquine for 4-5 years. She presented with a mottled, reticulated macular gray pigmentation involving the upper back and shoulders. Case 2 had a history of systemic lupus erythematosus and rheumatoid arthritis, treated with hydroxychloroquine for 1.5 years. She presented to the hospital for treatment of constrictive cardiomyopathy and was noted to have a blue macular pigmentation involving the right temple. The biopsies from both patients showed superficial dermal, yellow-brown, non-refractile and coarsely granular pigment deposition. A Fontana-Masson stain highlighted some of these granules, while the Perl's iron stain was negative. Rare, previous reports of hyperpigmentation indicate the presence of both melanin and hemosiderin in patients being treated with antimalarial medication. To our knowledge, this staining pattern for hydroxychloroquine has not been previously reported in the literature and supports that hydroxychloroquine, in addition to chloroquine, binds to melanin. PMID:18727667

Puri, Puja K; Lountzis, Nektarios I; Tyler, William; Ferringer, Tammie

2008-12-01

149

Evaluation of staining susceptibility of resin artificial teeth and stain removal efficacy of denture cleansers.  

PubMed

Abstract Objective. To assess the staining susceptibility of four acrylic resin (Ivostar, SR Vivodent PE, Major Dent, Integral) and a nanocomposite resin (Veracia) artificial teeth and to evaluate the stain removal efficacy of denture cleansers. Materials and methods. Sixty maxillary incisors of each brand (total = 300) were divided into three groups according to staining solution as coffee, red wine and tea. Baseline color measurements were performed with a spectrophotometer. Specimens were immersed in staining solutions for 14 h (2 h × 7 days) and then second color measurements were performed. Each group was further divided into four sub-groups according to denture cleanser as Corega tabs, Fittydent, NaOCl (0.5%) and distilled water (control) (n = 5). Specimens were immersed in denture cleansers for 8 h and third color measurements were made. Thus, the weekly simulation period was completed. This cycle was repeated 12 times to simulate a 3-month time period and measurements were performed at the end of the 4th, 8th and 12th cycles. ?E values were calculated and data were analyzed with 3-way repeated measures ANOVA and Bonferroni tests. Results. Significant color differences were found among the teeth and staining solutions, but all of the color differences were in the clinically acceptable range (?E < 5.5). Integral showed the highest ?E values for all solutions, while Ivostar and Vivodent demonstrated the lowest ?E values for red wine and tea solutions. There was no significant difference among the denture cleansers in terms of stain removal efficacy. Conclusions. Cross-linked acrylic (Integral) and nanocomposite (Veracia) resin teeth were more susceptible to staining. Denture cleansers were efficient on stain removal from artificial teeth. PMID:24807730

Kurtulmus-Yilmaz, Sevcan; Deniz, Sule Tugba

2014-11-01

150

The stain removal index (SRI): A new reflectometer method for measuring and reporting stain removal effectiveness  

Microsoft Academic Search

The development of laundry stain removal test methods is currently receiving attention in task groups of 3 major standards\\u000a developing organizations in the U.S. The need for such test methods is reflected in the proliferation of products for presoaking\\u000a or pretreatment of stained laundry items prior to washing or for addition to the main wash solution to help insure complete

O. W. Neiditch; K. L. Mills; G. Gladstone

1980-01-01

151

2, 2'-(phenylazanediyl) diacetic acid modified Fe3O4@PEI for selective removal of cadmium ions from blood  

NASA Astrophysics Data System (ADS)

A water-dispersible and supermagnetic nanocomposite (PAD-PEG-Fe3O4@PEI) has been successfully synthesized using polyethylenimine (PEI, Mol MW = 10000) coated supermagnetic Fe3O4-NH2 which was modified with 2, 2'-(phenylazanediyl) diacetic acid (PAD) through the bridge of poly(ethylene glycol) (PEG, Mol MW = 2000). The average particle size of PAD-PEG-Fe3O4@PEI was determined by TEM, and was about 50 nm. From magnetic hysteresis cycles for PAD-PEG-Fe3O4@PEI at room temperature, the saturation magnetization (Ms) was shown to be 58.14 emu g-1. Inductively coupled plasma spectrometry (ICP) analysis showed that the designed magnetic nanocomposite can remove 98% and 80% of Cd2+ from water and blood, respectively.A water-dispersible and supermagnetic nanocomposite (PAD-PEG-Fe3O4@PEI) has been successfully synthesized using polyethylenimine (PEI, Mol MW = 10000) coated supermagnetic Fe3O4-NH2 which was modified with 2, 2'-(phenylazanediyl) diacetic acid (PAD) through the bridge of poly(ethylene glycol) (PEG, Mol MW = 2000). The average particle size of PAD-PEG-Fe3O4@PEI was determined by TEM, and was about 50 nm. From magnetic hysteresis cycles for PAD-PEG-Fe3O4@PEI at room temperature, the saturation magnetization (Ms) was shown to be 58.14 emu g-1. Inductively coupled plasma spectrometry (ICP) analysis showed that the designed magnetic nanocomposite can remove 98% and 80% of Cd2+ from water and blood, respectively. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr11481j

Jin, Jun; Yang, Fang; Zhang, Fengwei; Hu, Wuquan; Sun, Shao-Bo; Ma, Jiantai

2012-01-01

152

Enhanced antimicrobial effects of combination of lactate and diacetate on Listeria monocytogenes and Salmonella spp. in beef bologna.  

PubMed

The antimicrobial activities of salts of organic acids such as lactate and acetate are well documented, but there is limited information on their effect when used in combination. We previously reported enhanced inhibition of Listeria monocvtogenes and Salmonella enterica serovar Enteritidis in sterile comminuted beef at 5 and 10 degrees C by combinations of sodium lactate (SL) (2.5%) and sodium diacetate (SDA) (0.2%). The present study was undertaken to evaluate the inhibitory effect of these salts, alone and in combination, in ready-to-eat (RTE) meat. Single strains and six-strain mixtures of each of the pathogens ( approximately 3 log CFU/g) were tested in beef bologna during aerobic storage at 5 and 10 degrees C for up to 60 days. The growth rate of the six-strain mixture of Listeria was faster than that of the single strain (Scott A) in the lactate/diacetate-free product. While each of the salts delayed growth of the listeriae at 5 degrees C, the effect of their combination was listericidal for the single strain and listeriostatic for the six-strain mixture. Enhanced inhibition by the salt combination was also observed at 10 degrees C. Salmonella numbers declined to undetectable levels in the untreated meat product and in each of the treatments after 20-30 days. However, the decline was more rapid in meat with the combination of the salts during storage at both 5 and 10 degrees C. Each of the salts further delayed the growth of the background microflora during storage at 5 degrees C, with their combinations showing the most effect. PMID:12051475

Mbandi, E; Shelef, L A

2002-06-25

153

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

NASA Astrophysics Data System (ADS)

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.

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

2013-11-01

154

Fluorescein angiography vs. optical coherence tomography for diagnosis of uveitic macular edema  

PubMed Central

Objective To evaluate agreement between fluorescein angiography (FA) and optical coherence tomography (OCT) for diagnosis of macular edema in patients with uveitis. Design Multicenter cross-sectional study Participants Four hundred seventy-nine eyes with uveitis of 255 patients Methods The macular status of dilated eyes with intermediate, posterior or panuveitis was assessed via Stratus-3 OCT and FA. Kappa statistics evaluated agreement between the diagnostic approaches. Main Outcome Measures Macular thickening (center point thickness ?240 ?m per reading center grading of OCT images-“MT”) and macular leakage (central subfield fluorescein leakage ?0.44 disk areas per reading center grading of FA images-“ML”); agreement amongst these outcomes in diagnosing “macular edema.” Results OCT (90.4%) more frequently returned usable information regarding macular edema than FA (77%) and biomicroscopy (76%). Agreement in diagnosis of MT and ML (?=0.44) was moderate. ML was present in 40% of cases free of MT, whereas MT was present in 34% of cases without ML. Biomicroscopic evaluation for macular edema failed to detect 40% and 45% of cases of MT and ML respectively and diagnosed 17% and 17% of cases with macular edema which did not have MT or ML respectively; these results may underestimate biomicroscopic errors (ophthalmologists were not explicitly masked to OCT and FA results). Among eyes free of ML, phakic eyes without cataract rarely (4%) had MT. No factors were found that effectively ruled out ML when MT was absent. Conclusion OCT and FA offered only moderate agreement regarding macular edema status in uveitis cases, probably because what they measure (MT and ML) are related but non-identical macular pathologies. Given its lower cost, greater safety, and greater likelihood of obtaining usable information, OCT may be the best initial test for evaluation of suspected macular edema. However, given that ML cannot be ruled out if MT is absent and vice versa, obtaining the second test after a negative result on the first seems justified when detection of ML or MT would alter management. Given that biomicroscopic evaluation for macular edema frequently erred, ancillary testing for macular edema seems indicated when knowledge of ML or MT status would affect management. PMID:23706700

Kempen, John H.; Sugar, Elizabeth A.; Jaffe, Glenn J.; Acharya, Nisha R.; Dunn, James P.; Elner, Susan G.; Lightman, Susan L.; Thorne, Jennifer E.; Vitale, Albert T.; Altaweel, Michael M.

2013-01-01

155

Wright-Giemsa and nonspecific esterase staining of cells.  

PubMed

This appendix provides two protocols for staining cells. The stains used in the Wright-Giemsa protocol, the Romanowsky stains, are a mixture of methylene blue (and other closely related thiazine dyes) and eosin. The staining protocol is a two-stage method that allows for a more intense staining of the nuclei than would be possible if the Wright and Giemsa stains were mixed together. In the second protocol, nonspecific esterase stain are used to identify cell types containing esterases that have a characteristic ability to split esters under particular conditions. In the staining method given here, the substrate, a-naphthyl butyrate, is incubated with cells under conditions in which esterases present in monocytes/macrophages split the substrate to yield an intermediate that can be coupled with a substance, hexazotized pararosaniline, to yield a colored precipitate. Thus, this staining reaction can be used to identify monocytes/macrophages in cell preparations. PMID:18432655

Strober, W

2001-05-01

156

Strategic Tool Use: Pencil Box Staining  

NSDL National Science Digital Library

This professional development video clip of students engaged in Common Core Practice Standard #5âUse 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.

Boston, Wghb

2013-01-01

157

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

PubMed Central

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

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

2003-01-01

158

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

PubMed

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

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

2014-07-01

159

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

PubMed Central

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

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

160

P53 detection by fluorescence lifetime on a hybrid fluorescein isothiocyanate gold nanosensor.  

PubMed

p53 is a tumor suppressor whose detection in the body is highly valuable as marker for early cancer diagnosis and prognosis. In this report we have studied constructs based on gold nanoparticles (NPs) functionalized with specific anti-p53 antibodies and with a fluoresceine derivative, FITC. The interaction of surface plasmons on gold NPs few nm in size, with fluorophores bound within few nanometers from the surface, induces changes in the fluorophore excited state lifetime. This parameter follows linearly the p53 concentration in solutions up to 200-400 pM, depending on the size of the NP, with an uncertainty approximatley =25 pM. We have evaluated the specificity of the nanosensor for p53 by testing it against bovine serum albumin, beta-lactoglobulin and lysozyme solutions. The titration of total cell extracts from p53 positive or p53-null cells with the anti-p53 antibody decorated gold NPs indicates that this construct is promising for possible applications to in vivo screening. PMID:20201230

Sironi, Laura; Freddi, Stefano; D'Alfonso, Laura; Collini, Maddalena; Gorletta, Tatiana; Soddu, Silvia; Chirico, Giuseppe

2009-12-01

161

Conformational Changes in the Intestinal Brush Border Sodium--Glucose Cotransporter Labeled with Fluorescein Isothiocyanate  

NASA Astrophysics Data System (ADS)

Fluorescein isothiocyanate (FITC) was used to label the rabbit intestinal brush border Na+-glucose carrier, identify the carrier protein on sodium dodecyl sulfate/polyacrylamide gel electrophoresis, and monitor the effect of ions and substrates on fluorescence quenching. Enriched brush border preparations were employed to study both glucose transport and FITC binding. FITC and a nonfluorescent analog (phenyl isothiocyanate, PITC) both inhibited Na+-dependent D-glucose transport irreversibly. Inhibition was blocked completely by the presence of Na+ and D-glucose during labeling. PITC was used to label nonspecific amino groups in the presence of glucose and Na+, and then the glucose carrier was labeled with FITC in the absence of substrates. Fluorescence of FITC bound to the carrier was quenched specifically with Na+ in a saturable fashion, and this indicates a Na+-dependent conformational change in the carrier. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis of FITC-labeled membranes revealed specific labeling of a 71,000-dalton peptide. We conclude that Na+ induces a conformational shift in the 71,000-dalton glucose carrier, and this is quite consistent with the kinetics of Na+-depedent glucose transport in these membranes.

Peerce, Brian E.; Wright, Ernest M.

1984-04-01

162

Electrical conductivity and dielectric relaxation behavior of fluorescein sodium salt (FSS)  

NASA Astrophysics Data System (ADS)

The ac and dc electrical conductivities and dielectric properties of the fluorescein sodium salt (FSS) have been investigated. The direct current (dc) conductivity shows that the compound is a typical organic semiconductor, as its electrical conductivity increases with increasing temperature. The dc electrical activation energy ?E? and the room temperature electrical conductivity equal 0.35 eV and 1.26×10-9 (, respectively. The alternating current (ac) conductivity of the investigated compound obeys the power law : ?(?)=A(T).?, where s<1. The obtained results have been discussed in terms of the correlated barrier hopping (CBH) model. The density of localized states NF(E) at the Fermi level and the binding energy Wm were calculated. The dielectric constant ?1(?) and dielectric loss ?2(?) have been found to decrease with increasing frequency and to increase with increasing temperature over the studied ranges. The value of the maximum barrier height WM obtained from Austin and Guintini equations agree with each other. The correlation between the ac conduction and the dielectric properties in organic FSS were verified.

Mansour, Sh. A.; Yahia, I. S.; Sakr, G. B.

2010-08-01

163

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

PubMed Central

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

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

2002-01-01

164

Selective Hg(II) Detection in Aqueous Solution with Thiol Derivatized Fluoresceins  

PubMed Central

The syntheses and photophysical properties of MS2 and MS3, two asymmetrically derivatized fluorescein-based dyes designed for Hg(II) detection, are described. These sensors each contain a single pyridyl-amine-thiol metal-binding moiety, form 1:1 complexes with Hg(II), and exhibit selectivity for Hg(II) over other Group 12 metals, alkali and alkaline earth metals, and most divalent first-row transition metals. Both dyes display superior brightness (? × ?) and fluorescence enhancement following Hg(II) coordination in aqueous solution. At neutral pH, the fluorescence turn-on derives from greater brightness due to increased molar absorptivity. At higher pH, photoinduced electron transfer (PET) quenching of the free dye is enhanced, and the Hg(II)-induced turn-on also benefits from alleviation of this pathway. MS2 can detect ppb levels of Hg(II) in aqueous solution, demonstrating its ability to identify environmentally relevant concentrations of Hg(II). PMID:16529499

Nolan, Elizabeth M.; Racine, Maryann E.; Lippard, Stephen J.

2008-01-01

165

Fluorescein hydrazones as novel nonintercalative topoisomerase catalytic inhibitors with low DNA toxicity.  

PubMed

Fluorescein hydrazones (3a-3l) were synthesized in three steps with 86-91% overall yields. Topo I- and II?-mediated relaxation and cell viability assay were evaluated. 3d inhibited 47% Topo I (camptothecin, 34%) and 20% Topo II (etoposide 24%) at 20 ?M. 3l inhibited 61% Topo II (etoposide 24%) at 20 ?M. 3d and 3l were further evaluated to determine their mode of action with diverse methods of kDNA decatenation, DNA-Topo cleavage complex, comet, DNA intercalating/unwinding, and Topo II?-mediated ATP hydrolysis assays. 3d functioned as a nonintercalative dual inhibitor against the catalytic activities of Topo I and Topo II?. 3l acted as a Topo II? specific nonintercalative catalytic inhibitor. 3d activated apoptotic proteins as it increased the level of cleaved capase-3 and cleaved PARP in a dose- and time-dependent manner. The dose- and time-dependent increase of G1 phase population was observed by treatment of 3d along with the increase of p27(kip1) and the decrease of cyclin D1 expression. PMID:25333701

Rahman, A F M Motiur; Park, So-Eun; Kadi, Adnan A; Kwon, Youngjoo

2014-11-13

166

Staining etched epoxy resin sections for light microscopy.  

PubMed

Staining of etched sections for light microscopy is described. Azan staining was successful after treatment with potassium dichromate and the use of concentrated dye solutions. To remove osmium for hematoxylin-eosin staining, removal by reduction with ferrocene was used instead of oxidation. Highly selective differentiation after hematoxylin staining was achieved using p-toluenesulfonic acid-DMSO. To enhance eosin staining, a 2-bromoethylamine link between eosin and the tissue was used. Ferrocene also facilitated counterstaining of nuclei with hematoxylin after the PAS reaction. Periodic acid-methenamine silver staining was carried out without modification. PMID:7578588

Iwadare, T; Arai, T

1995-03-01

167

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

MedlinePLUS

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

168

Laser therapy in plastic surgery: decolorization in port wine stains  

NASA Astrophysics Data System (ADS)

For the first time laserotherapy is described as a method of port wine stain decolorization in plastic surgery. The authors present their 20-year experience in the treatment of port wine stains with the argon laser and dye laser.

Peszynski-Drews, Cezary; Wolf, Leszek

1996-03-01

169

Wright-Giemsa and nonspecific esterase staining of cells.  

PubMed

This appendix provides two protocols for staining cells. The stains used in the Wright-Giemsa protocol, the Romanowsky stains, are a mixture of methylene blue (and other closely related thiazine dyes) and eosin. The staining protocol is a two-stage method that allows for a more intense staining of the nuclei than would be possible if the Wright and Giemsa stains were mixed together. In the second protocol, nonspecific esterase stain are used to identify cell types containing esterases that have a characteris termediate that can be coupled with a substance, hexazotized pararosaniline, to yield a colored precipitate. Thus, this staining reaction can be used to identify monocytes/macrophages in cell preparations. PMID:18770656

Strober, W

2001-05-01

170

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2012 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

2012-04-01

171

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2013 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

2013-04-01

172

21 CFR 864.1850 - Dye and chemical solution stains.  

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

2014-04-01

173

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2011 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

2011-04-01

174

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2010 CFR

... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...

2010-04-01

175

Optimization of a rapid test by using fluorescein-conjugated monoclonal antibodies for detection of Chlamydia trachomatis in clinical specimens.  

PubMed Central

A mixture of two fluorescein isothiocyanate-conjugated monoclonal antibodies (MAbs) was used to optimize a direct specimen test (Chlamydia Direct Specimen Test IF; Clonatec, Paris, France) for detection of chlamydial elementary bodies in clinical specimens. One MAb reacted with a subspecies-specific epitope of the major outer membrane protein (molecular weight 43,000) of Chlamydia trachomatis, whereas the other reacted with the periodate-sensitive genus-specific antigen (molecular weight 11,000) of Chlamydia spp. Nonfat dry milk was the most efficient additive at suppressing the fluorescent background and was included in the antibody preparation. Fc-dependent binding of fluorescein-conjugated MAbs to protein A-containing Staphylococcus aureus was inhibited by addition of purified rabbit immunoglobulin. The Chlamydia Direct Specimen Test IF was compared with tissue culture isolation by using 309 genital specimens. The sensitivity and specificity were 77.4 and 98%, respectively. Images PMID:2449456

Pouletty, P; Martin, J; Catalan, F; Garcia-Gonzalez, M; Morellet, I; Bettinger, S; Kadouche, J

1988-01-01

176

Temperature and pH triggered release characteristics of water/fluorescein from 1-ethyl-3-methylimidazolium ethylsulfate based ionogels.  

PubMed

A crosslinked poly(N-isopropylacrylamide) ionogel encapsulating an ionic liquid exhibits improved transmittance properties, enhanced water uptake/release, greater thermal actuation behaviour and distinct solvatomorphology over its hydrogel equivalent. It was also found that the rate of release of fluorescein pre-loaded into membranes was considerably enhanced for ionogels compared to equivalent hydrogels, and could be triggered through changes in pH and temperature. PMID:23579593

Gallagher, Simon; Kavanagh, Andrew; Florea, Larisa; MacFarlane, Douglas R; Fraser, Kevin J; Diamond, Dermot

2013-05-21

177

Non-photorealistic Rendering of Images as Evolutionary Stained Glass  

E-print Network

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

Ashlock, Dan

178

bleachingIn vitro chemical stain removal by 'whitening' toothpastes  

Microsoft Academic Search

Background A considerable number of toothpastes are available as tooth whitening products. Most appear to contain ingredients that might remove extrinsic stains rather than change natural tooth colour. Extrinsic stain removal could be achieved by physical or chemical means.Aim The purpose of this study was to measure the chemical stain removal properties of a range of whitening toothpaste products and

Diana Scarrott

2000-01-01

179

Indocyanine green selectively stains the internal limiting membrane  

Microsoft Academic Search

PURPOSE: To demonstrate whether indocyanine green stains the inner limiting membrane of the retina or residual vitreous cortex.METHODS: We report on the intraoperative staining patterns of the vitreomacular interface in 10 eyes of 10 consecutive patients who underwent vitrectomy with indocyanine green staining for macular hole formation and diffuse diabetic macular edema.RESULTS: In five eyes of five patients with macular

Arnd Gandorfer; Elisabeth M. Messmer; Michael W. Ulbig; Anselm Kampik

2001-01-01

180

A Chemical Stain for Identifying Arsenic-Treated Wood  

E-print Network

.0 kg/m3 CCA-Treated Wood Figure II.7 Picture of Stannous Chloride Stain (left) and Diluted Stannous Chloride Stain (right) Figure II.8 Picture of PAN Indicator and Diluted Stannous Chloride Stain on New Untreated, Borate, ACQ, CBA, and 4.0 kg/m3 CCA-Treated Wood Figure II.9 Picture of Wipe Samples from

Florida, University of

181

Cigarette staining and cleaning of a maxillofacial silicone  

SciTech Connect

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.

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

1983-07-01

182

Lower limits of fluorescein and indocyanine green dye for digital cSLO fluorescence angiography  

PubMed Central

Background: With the advent of digital confocal scanning laser ophthalmoscopy it is possible to detect low levels of fluorescence. Here we used a novel confocal scanning laser ophthalmoscope (cSLO) to determine lower limits of dye required for fluorescein (FL) and indocyanine green (ICG) angiography. Methods: A cSLO (Heidelberg retina angiograph 2, Heidelberg Engineering, Dossenheim, Germany) with an optically pumped solid state laser (488 nm) for FL and a diode laser (790 nm) for ICG angiography (FL/ICG-A) was used. 62 FL-As were performed in 53 patients and 45 ICG-As were performed in 39 patients with neovascular age related macular degeneration. The volume and overall dye content of bolus injections was gradually tapered (FL: 500 mg, 250 mg, 200 mg, 166 mg, 100 mg; ICG: 25 mg, 20 mg, 15 mg, 10 mg, 5 mg, 2.5 mg), while dye concentrations were kept constant at 100 mg/ml for FL and at 5 mg/ml for ICG. Images were obtained 1, 5, 15, and 30 minutes after dye injection. Image quality was evaluated by two independent readers using standardised criteria. Results: For amounts down to 166 mg for FL and to 5 mg for ICG, sufficient image quality was achieved during all phases following injection. Only late phase images showed less contrast compared to typically used dye amounts, which was irrelevant for interpretation and clinical management. Conclusions: With the increased sensitivity of this novel cSLO system, amounts of injected dye during FL-A can be reduced to one third for FL and to one fifth for ICG without relevant loss of image quality or information compared to conventionally used dye levels. These amounts can be used for routine angiography and allow relevant savings for units performing FL-A. PMID:16299141

Bindewald, A; Stuhrmann, O; Roth, F; Schmitz-Valckenberg, S; Helb, H-M; Wegener, A; Eter, N; Holz, F G

2005-01-01

183

Cannabinoid 1 receptors in keratinocytes attenuate fluorescein isothiocyanate-induced mouse atopic-like dermatitis.  

PubMed

Atopic dermatitis is a chronic inflammatory disease characterized by an impaired epidermal barrier function combined with a chronic Th2-type inflammatory response and an intense pruritus. Here, we used an experimental mouse model for Th2-type contact hypersensitivity (CHS) to fluorescein isothiocyanate (FITC) to investigate the potential role of cannabinoid 1 receptors (CB1) in the pathophysiology of mouse atopic-like dermatitis. Mice lacking CB1 receptors globally (Cnr1(-/-) ) or specifically in keratinocytes (KC-Cnr1(-/-) ) as well as wild-type (WT) control mice were sensitized and challenged with FITC. We examined ear swelling responses, transepidermal water loss, Th2-type skin inflammatory responses and serum IgE levels. Both Cnr1(-/-) and KC-Cnr1(-/-) showed enhanced CHS responses to FITC and a delayed epidermal barrier repair when compared with WT mice. mRNA levels for IL-4, thymic stromal lymphopoietin (TSLP) and CCL8, as well as eosinophil activity, were significantly increased in inflamed ear tissue of FITC-challenged Cnr1(-/-) and KC-Cnr1(-/-) mice. Importantly, CB1 receptor-deficient keratinocytes secreted increased levels of TSLP, a proinflammatory mediator that drives Th2-type skin inflammation in atopic dermatitis, under basal and Th2-type inflammatory conditions. Taken together, our results demonstrate that CB1 receptors in keratinocytes help to maintain epidermal barrier homoeostasis and attenuate Th2-type allergic inflammatory responses. Based on our work, we propose that enhanced epidermal allergen penetrance cooperates with increased production of TSLP and CCL8 by epidermal keratinocytes for the induction of type 2 CD4+ T helper cells. Our results place keratinocytes at the cross-roads of outside-in and inside-out pathophysiologic mechanisms of atopic dermatitis. PMID:24750433

Gaffal, Evelyn; Glodde, Nicole; Jakobs, Mira; Bald, Tobias; Tüting, Thomas

2014-06-01

184

Detection of Cystoid Macular Edema with Three-Dimensional Optical Coherence Tomography versus Fluorescein Angiography  

PubMed Central

Purpose. To compare the sensitivity and reproducibility of three-dimensional optical coherence tomography (3D-OCT) and fluorescein angiography (FA) for the detection of cystoid macular edema (CME). Methods. Data were retrospectively collected from all patients who underwent digital FA and 512 × 128 horizontal raster 3D-OCT scans on the same day in a retina subspecialty clinic. Images were reviewed independently by four reading center graders and adjudicated as a group to render a single result for each eye and each imaging modality. The ? statistic was used to determine the level of agreement between graders for each modality. The sensitivity of each imaging modality for CME detection was calculated by using the presence of CME on either modality as the ground truth; subgroup analysis was performed according to disease diagnosis and lens status. Results. Four hundred thirteen eyes of 207 patients were included in the analysis. Intergrader agreement was higher for 3D-OCT than for FA both before (?OCT = 0.61, ?FA = 0.43) and after adjudication (?OCT = 0.74, ?FA = 0.58).The sensitivity for detection of definite CME was higher for 3D-OCT (95%, 144/151 cases) than for FA (44%, 67/151 cases). Definite FA (+) 3D-OCT (?) CME was identified in 1 eye (0.2%), whereas definite FA (?) 3D-OCT (+) CME was identified in 40 eyes (10%). No significant associations between CME detection and lens examination or disease diagnosis were observed. Conclusions. In this study, 3D-OCT was more sensitive and had better intergrader agreement than did FA for the detection of CME. PMID:20357195

Ouyang, Yanling; Keane, Pearse A.; Sadda, Srinivas R.

2010-01-01

185

Ultrafast tissue staining with chemical tags  

PubMed Central

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

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

2014-01-01

186

Estrogen staining in breast carcinoma by PAP methods compared to CEA and ferritin staining.  

PubMed

The aims of this paper are to demonstrate the stainability of estrogen, CEA, and ferritin in breast carcinomas, fibroadenomas, and fibrocystic diseases; to examine whether the findings of endogenous estrogen using the immunohistochemical detection method are related to estrogen receptor (ER) assays; and to determine whether the stainability of estrogen, CEA, and ferritin were related to the prognosis of breast carcinomas. In breast cancer, the stainability of estrogen using the peroxidase-antiperoxidase (PAP) method was positively correlated with the dextran-coated charcoal (DCC) assay for ER. In breast cancers, the percentage of positive staining was 46% for estrogen, 48% for CEA, and 47% for ferritin. With all three stains, significant differences were observed between cancer and benign diseases. Cases that were both positive for estrogen staining and negative for CEA showed a good prognosis after the recurrence of disease. Our data suggest that the immunohistochemical staining of estrogen, CEA, and ferritin might predict the biological behavior of breast carcinomas and be a prognostically useful indicator of breast cancer patients. PMID:2436774

Osamu, K; Takashi, M; Yohichi, T; Yasuo, U; Tetsuro, Y; Yoshiro, F; Toshio, T

1987-01-01

187

Asymmetric catalysis, part 91: Enantioselective monophenylation of cis -1,2-cyclopentanediol with triphenylbismuth diacetate and chiral copper(II) complexes as catalysts  

Microsoft Academic Search

The monophenylation ofcis-1,2-cyclopentanediol with triphenylbismuth diacetate in the presence of chiral Cu(II) complexes as catalysts gavecis-2-hydroxy-1-phenoxy-cyclopentane with enantiomeric excesses up to 38%. The optically active ligands used were triamine derivatives of 2,6-bis(aminomethyl)pyridine and diamine derivatives of 2-(aminomethyl)pyridine. Selectivity in the monophenylation occurred only in the presence of the latter as auxiliary ligands.

H. Brunner; T. Chuard

1994-01-01

188

Comparison of three staining methods for detecting microsporidia in fluids.  

PubMed Central

Calcofluor white 2MR, modified trichrome blue, and indirect immunofluorescent antibody (IFA) staining methods were evaluated and compared for detecting microsporidia in stool. Serial 10-fold dilutions of Encephalitozoon (Septata) intestinalis were prepared in three formalinized stool specimens or in Tris-buffered saline. Ten-microliter aliquots were smeared onto glass slides, fixed with methanol, stained, and read by at least three individuals. The results indicated that the calcofluor stain was the most sensitive method, required approximately 15 min to perform, but did generate some false-positive results due to similarly staining small yeast cells. The modified trichrome blue stain was nearly as sensitive as the calcofluor stain and allowed for easier distinction between microsporidia and yeast cells. This stain, however, required approximately 60 min to perform. The IFA stain with polyclonal murine antiserum against E. intestinalis was the least sensitive of the methods and required approximately 130 min to perform. The lower limit of detection with the calcofluor and modified trichrome stains was a concentration of about 500 organisms in 10 microliters of stool to detect one microsporidian after viewing 50 fields at a final magnification of x1,000. Reliability was also addressed by use of 74 stool, urine, and intestinal fluid specimens, 50 of which were confirmed for the presence of microsporidia by transmission electron microscopy (TEM). All TEM-positive specimens were detected by calcofluor and modified trichrome blue staining. Ten specimens were not detected by the IFA stain. An additional seven TEM-negative specimens were read positive for microsporidia with the calcofluor stain, and of these, five also were read positive with the modified trichrome blue stain. The resulting diagnostic paradigm was to screen specimens with the calcofluor stain and to confirm the results with the modified trichrome stain. IFA, which was less sensitive, may become useful for microsporidian species identification as specific antibodies become available. PMID:8586689

Didier, E S; Orenstein, J M; Aldras, A; Bertucci, D; Rogers, L B; Janney, F A

1995-01-01

189

Automated single-slide staining device. [in clinical bacteriology  

NASA Technical Reports Server (NTRS)

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

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

1975-01-01

190

Centrifuge-operated specimen staining method and apparatus  

NASA Technical Reports Server (NTRS)

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.

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

1999-01-01

191

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

SciTech Connect

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

Rush, J.D.; Maskos, Z. (Louisiana State Univ., Baton Rouge (USA))

1990-03-07

192

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

PubMed Central

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

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

2009-01-01

193

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

PubMed Central

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

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

2011-01-01

194

Highly sensitive and selective fluorimetric methods for the determination of iron(III) and manganese(II) using fluorescein\\/hydrogen peroxide\\/triethylenetetramine and fluorescein\\/hydrogen peroxide\\/triethylenetetramine\\/tiron, respectively  

Microsoft Academic Search

Highly sensitive and selective spectrofluoriphotometric determinations of iron(III) with fluorescein(Fl)-hydrogen peroxide-triethylenetetramine (TETA), and manganese(II) with Fl-hydrogen peroxide-TETA-tiron are proposed. The methods are based on the inhibition of the oxidizing decomposition of Fl-hydrogen peroxide solution in the presence of iron(III)-TETA or manganese(II)-TETA-tiron combination. The calibration graphs are linear in the ranges of up to 220 ng iron(III) and up to 270

I. Mori; Y. Fujita; K. Ikuta; Y. Nakahashi; K. Kato

1989-01-01

195

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

PubMed

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

Biesemeier, Antje; Schraermeyer, Ulrich; Eibl, Oliver

2011-07-01

196

Determining an Appropriate Method to Simulate Pump Shear on the Diatom Nitzschia sp. and a Methodology to Quantify the Effects  

E-print Network

C.. ............................................................................... 19 10 An example of corresponding images taken under brightfield and fluorescent light with labels and notations for data acquisition. ................ 22 11 Graph of cell viability as determined by FDA stain and image analysis... was quantified using a staining procedure utilizing fluorescein diacetate (FDA) to stain only the healthy cells. A stock solution of 46 mg of FDA stain mixed with 10 mL acetone was added to an algae solution at a ratio of 10 ?l of FDA solution to 1 ml of algae...

Lassig, Jarrett

2012-12-13

197

Visible luminescence from silicon wafers subjected to stain etches  

NASA Technical Reports Server (NTRS)

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.

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

1992-01-01

198

Dissociation kinetics of 1,7-diaza-4,10,13-trioxacyclopentadecane-N,N'-diacetic acid complexes of lanthanides  

SciTech Connect

The dissociation kinetics of 1,7-diaza-4,10,13-trioxacyclopentadecane-N,N'-diacetic acid (K21DA) complexes of lanthanide(III) ions were studied in acetate-acetic acid buffer medium, over the acid concentration range of 8.4 x 10/sup -6/-2.5 x 10/sup -4/ M and at a constant ionic strength of 0.1 M (LiClO/sub 4/). Copper(II) was used as the scavenger of free ligand, and the rates of dissociation of these complexes have been found to be independent of (Cu/sup 2 +/). All the complexes exhibit acid-independent and acid-dependent pathways. Lighter lanthanide complexes display a first-order dependence upon (H/sup +/) in the pH range studied. The complexes of heavier lanthanides show (H/sup +/) dependence at low acid concentrations but become acid-independent at high acid concentrations. Influence of acetate content in the buffer and total electrolyte concentration on the rate of dissociation has also been investigated. The observed rate constants for erbium, ytterbium, and lutetium complexes do not show a significant dependence on acetate concentration, but lanthanum and europium complexes do exhibit a first-order dependence on (acetate). All the complexes under study respond similarly with change in electrolyte concentration; i.e., the rate constants decrease with increase in (electrolyte). Activation parameters for both self-dissociation and acid-catalyzed dissociation pathways have been obtained for lanthanum, europium, erbium, and lutetium complexes, from the temperature dependence of rate constants in the 15-45 /sup 0/C range. The results are compared with those of the lanthanide-polyamino polycarboxylate systems, and possible mechanisms are discussed. 41 references, 4 figures, 6 tables.

Sekhar, V.C.; Chang, C.A.

1986-06-04

199

Enhanced inhibition of Listeria monocytogenes and salmonella enteritidis in meat by combinations of sodium lactate and diacetate.  

PubMed

The antimicrobial activities of sodium lactate (SL) and sodium acetate (SA) are well documented, but there is limited information on the effect of their combination or of the combination of SL and sodium diacetate (SDA) on survival and growth of Listeria monocytogenes and salmonellae in meat. Effects of SL (1.8 and 2.5%), SDA (0.1 and 0.2%), or SA (0.2%) and their combinations on the behavior of L monocytogenes and Salmonella enterica serovar Enteritidis were investigated in sterile comminuted beef (pH 6.3, 79% moisture) during storage at 5 and 10 degrees C. Although L. monocytogenes grew faster than Salmonella Enteritidis in control samples at 10 degrees C, numbers of both pathogens increased from 3.5 to approximately 8.0 log CFU/g after 20 days. SL (1.8%) decreased the growth rate of both L. monocytogenes and Salmonella Enteritidis. SDA (0.2%) was more effective than SL in decreasing the growth rate of L monocytogenes, and it caused a more than 1 log CFU/g decline in initial numbers of Salmonella Enteritidis during storage for 25 days at 10 degrees C. Synergy was observed by combinations of SL and SDA. Combinations of 2.5% SL and 0.2% SDA were bacteriostatic to L. monocytogenes and bactericidal to Salmonella Enteritidis after 20 days at 10 degrees C. At 5 degrees C, a listeriostatic effect was produced by 1.8% SL + 0.1% SDA, whereas numbers of Salmonella Enteritidis were less than 10 cells/g after refrigeration for 30 days. Although SA was consistently and significantly less inhibitory than SDA, its mixtures with SL also demonstrated synergistic activity against both pathogens. Combinations of 2.5% SL and 0.2% SDA can be expected to greatly enhance the safety of refrigerated and temperature-abused ready-to-eat meats. PMID:11347993

Mbandi, E; Shelef, L A

2001-05-01

200

Vascular invasion of colorectal carcinoma readily visible with certain stains  

Microsoft Academic Search

We made use of hematoxylin and eosin (H&E) stain, Verhoeff van-Gieson stain for elastic tissue (EVG), and factor VIII-related antigen (FVIII-RA) to stain tissues excised from 94 patients with colorectal carcinoma. Of these 94, 49 died of disease within two years (Group I), and 45 survived for five years or longer (Group II) after surgery. In the tissues from both

Tetsuya Inoue; Masaki Mori; Reishi Shimono; Hiroyuki Kuwano; Keizo Sugimachi

1992-01-01

201

The blood-brain barrier penetration and distribution of PEGylated fluorescein-doped magnetic silica nanoparticles in rat brain  

SciTech Connect

PEGylated PAMAM conjugated fluorescein-doped magnetic silica nanoparticles (PEGylated PFMSNs) have been synthesized for evaluating their ability across the blood-brain barrier (BBB) and distribution in rat brain. The obtained nanoparticles were characterized by transmission electron microscopy (TEM), thermal gravimetry analyses (TGA), zeta potential ({zeta}-potential) titration, and X-ray photoelectron spectroscopy (XPS). The BBB penetration and distribution of PEGylated PFMSNs and FMSNs in rat brain were investigated not only at the cellular level with Confocal laser scanning microscopy (CLSM), but also at the subcellular level with transmission electron microscopy (TEM). The results provide direct evidents that PEGylated PFMSNs could penetrate the BBB and spread into the brain parenchyma.

Ku, Shuting [School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing (China)] [School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing (China); Yan, Feng [Xuanwu Hospital, Capital Medical University, Beijing (China)] [Xuanwu Hospital, Capital Medical University, Beijing (China); Wang, Ying [Basic Medical Sciences, Capital Medical University, Beijing (China)] [Basic Medical Sciences, Capital Medical University, Beijing (China); Sun, Yilin [Beijing Neurosurgical Institution, Beijing (China)] [Beijing Neurosurgical Institution, Beijing (China); Yang, Nan [Basic Medical Sciences, Capital Medical University, Beijing (China)] [Basic Medical Sciences, Capital Medical University, Beijing (China); Ye, Ling, E-mail: lingye@ccmu.edu.cn [School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing (China)] [School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing (China)

2010-04-16

202

Asymmetric incorporation of (/sup 14/C)cyanate and of fluorescein isothiocyanate in mamillary body of conditioned rats  

SciTech Connect

A marked decrease in overall learning capacity has been observed in rats injected with cyanate. Therefore it was of interest to test whether learning influenced carbamylation of brain proteins. Incorporation of (/sup 14/C)cyanate into proteins of the mamillary body was selectively modified following operant conditioning of the rat, so that trained rats showed an asymmetric image with higher levels of incorporation in the right side than in the left side, as compared to control rats. These results were confirmed using fluorescein isothiocyanate. The asymmetry persisted once the learning had been well established.

Burgal, M.; Montes, F.; Grisolia, S.

1988-05-01

203

The crystallization kinetics and thermal conductivity of alumina/fluorescein sodium salt (Al2O3/FSS) composites  

NASA Astrophysics Data System (ADS)

The thermal conductivity and crystallization mechanism of alumina (Al2O3)/fluorescein sodium salt (FSS) composites prepared by the powder metallurgy method have been investigated by means of differential thermal analysis. The Kissinger method is applied to determine the crystallization kinetics from the endotherm peaks. The activation energy E and Avrami parameter n were calculated. The kinetic parameters (E and n) have made it possible to postulate the type of crystal growth exhibited in the crystallization process. The crystallization growth is found to be one-dimensional for the composite system. The thermal conductivity of the composite system was also determined by differential scanning calorimetry.

Yakuphanoglu, Fahrettin; Sekerci, M.

2005-01-01

204

Capillary zone electrophoresis separation and laser-induced fluorescence detection of zeptomole quantities of fluorescein thiohydantoin derivatives of amino acids.  

PubMed

Capillary zone electrophoresis, when combined with laser-induced fluorescence, is a very powerful technique for the separation and determination of minute amounts of labeled amino acids. This paper presents the determination of the fluorescein thiohydantoin derivative of 17 amino acids which takes 13.5 min. The detector, based on laser-induced fluorescence, is optimized with respect to laser power to produce detection limits, three standard deviations above background, ranging from 1 to 2 zeptomoles (1 zeptomole = 1 zmole = 10(-21) = 600 analyte molecules) injected onto the capillary. PMID:18965357

Wu, S; Dovichi, N J

1992-02-01

205

Stain defect detection for mobile phone camera modules  

NASA Astrophysics Data System (ADS)

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.

Hong, Sehee; Lee, Chulhee

2014-03-01

206

Dynamic staining of Bacillus endospores with Thioflavin T.  

PubMed

Rapid detection and identification of endospores presents a range of complex challenges. Dynamic staining approach, developed in our lab, utilizes the time-course fluorescence enhancement of an amyloid-staining dye, Thioflavin T (ThT), after mixing with intact endospores. We examined the kinetics of staining Bacillus atrophaeus and Bacillus thuringiensis endospores, and the rates of staining were different for the two bacilli when intact endospores were treated with ThT. This finding demonstrates an avenue for attaining information about the sporulated bacterial species without lysing, germinating or other pretreatment steps. PMID:23365938

Upadhyayula, Srigokul; Lam, Samuel; Ha, Alice; Malik-Chaudhry, Harbani K; Vullev, Valentine I

2012-01-01

207

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

PubMed

Abstract 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

Belaldavar, C; Hallikerimath, S; Angadi, Pv; Kale, Ad

2014-11-01

208

The Transcriptional Response of Listeria monocytogenes during Adaptation to Growth on Lactate and Diacetate Includes Synergistic Changes That Increase Fermentative Acetoin Production?†  

PubMed Central

The organic acids lactate and diacetate are commonly used in combination in ready-to-eat foods because they show synergistic ability to inhibit the growth of Listeria monocytogenes. Full-genome microarrays were used to investigate the synergistic transcriptomic responses of two L. monocytogenes strains, H7858 (serotype 4b) and F6854 (serotype 1/2a), to these two organic acids under conditions representing osmotic and cold stress encountered in foods. Strains were exposed to brain heart infusion (BHI) broth at 7°C with 4.65% water-phase (w.p.) NaCl at pH 6.1 with (i) 2% w.p. potassium lactate, (ii) 0.14% w.p. sodium diacetate, (iii) the combination of both at the same levels, or (iv) no organic acids as a control. RNA was extracted 8 h after exposure, during lag phase, to capture gene transcription changes during adaptation to the organic acid stress. Significant differential transcription of 1,041 genes in H7858 and 640 genes in F6854 was observed in at least one pair of the 4 different treatments. The effects of combined treatment with lactate and diacetate included (i) synergistic transcription differences for 474 and 209 genes in H7858 and F6854, respectively, (ii) differential transcription of genes encoding cation transporters and ABC transporters of metals, and (iii) altered metabolism, including induction of a nutrient-limiting stress response, reduction of menaquinone biosynthesis, and a shift from fermentative production of acetate and lactate to energetically less favorable, neutral acetoin. These data suggest that additional treatments that interfere with cellular energy generation processes could more efficiently inhibit the growth of L. monocytogenes. PMID:21666015

Stasiewicz, Matthew J.; Wiedmann, Martin; Bergholz, Teresa M.

2011-01-01

209

Development of rapid staining protocols for laser-capture microdissection of brain vessels from human and rat coupled to gene expression analyses.  

PubMed

Laser-capture microdissection (LCM) is a technique that enables selective extraction of desired cells from heterogeneous tissues compatible with subsequent molecular analyses. The specific visualization of desired cell types prior to LCM is essential for achieving selective capture. We have developed rapid and selective staining protocols for LCM extraction of microvessels from human and rat brain. Vessels in human and rat brain sections were visualized by a 2 min exposure to fluorescein-labeled lectins Ulex Europeaus Agglutinin I (UEA I) and Ricinus Communis Agglutinin I (RCA I), respectively. Immunohistochemical staining for the endothelial-specific marker, Factor VIII-related antigen (FVIII-rAg), co-localized with that for either UEA I or RCA I, confirming the selective staining of vascular structures with these lectins. Both brain vessels and perivascular parenchyma were captured using LCM, followed by RNA isolation. RT-PCR analyses demonstrated the enrichment of LCM-captured vessels and parenchyma in FVIII-rAg and GFAP mRNA, respectively. LCM-captured human vessels also expressed the tight junction-specific gene, zonula occludens 1 (ZO-1). LCM extraction of vessels from brain sections can be used to perform molecular fingerprinting of neurovascular unit in various brain pathologies. PMID:14757343

Mojsilovic-Petrovic, Jelena; Nesic, Momir; Pen, Ally; Zhang, Wandong; Stanimirovic, Danica

2004-02-15

210

catena-Poly[[[diaqua-copper(II)]-?-2,2'-{[p-phenyl-enebis(oxymethyl-ene)]bis-(pyridinium-3,1-di-yl)}diacetate] dibromide].  

PubMed

The title centrosymmetric coordination polymer, {[Cu(C(22)H(20)N(2)O(6))(H(2)O)(2)]Br(2)}(n), formed by the reaction of the flexible double betaine ligand 2,2'-{[p-phenyl-enebis(oxymethyl-ene)]bis-(pyridine-3,1-di-yl)}diacetic acid with CuBr(2), contains a Cu(II) atom ( symmetry) which is surrounded by two water molecules and bridged by two anions in a square-planar coordination. In the crystal, polymeric zigzag chains are linked via O-H?Br inter-actions, forming a two-dimensional network extending parallel to (011). PMID:21579269

Pan, Wei-Cheng; Lian, Hong-Lei

2010-01-01

211

7 CFR 3201.87 - Wood and concrete stains.  

...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...

2014-01-01

212

7 CFR 3201.87 - Wood and concrete stains.  

Code of Federal Regulations, 2013 CFR

...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...

2013-01-01

213

Black stain and dental caries in Filipino schoolchildren  

Microsoft Academic Search

Black stain is defined as dark pigmented exogenous substance in lines or dots parallel to the gingival margin and firmly adherent to the enamel at the cervical third of the tooth crowns in the primary and permanent dentition. OBJECTIVES: This study was conducted to assess the prevalence of black stain on teeth of Filipino children and to determine a possible

Roswitha Heinrich-Weltzien; Bella Monse; Wim van Palenstein Helderman

2009-01-01

214

Effects of metabolic inhibitors on vital staining with methylene blue  

Microsoft Academic Search

The peripheral innervation of murine skin was vitally stained with methylene blue and the effects of various metabolic inhibitors on the staining process were investigated. The observed effects suggest that uptake of the dye is dependent on the integrity of a membrane-associated adenosine triphosphatase and an unidentified metallo-enzyme. Glycolysis, the citric acid cycle and the cytochrome system are not necessary

J. A. Kiernan

1974-01-01

215

Removing foxing stains from old paper at 157nm  

Microsoft Academic Search

Using a molecular fluorine laser at 157 nm foxing stains were removed successfully from a 16th century old paper. Laser cleaning of stains and foxing from old paper manuscripts is far more effective at 157 nm in comparison to different wavelengths without any yellowish after-effect on the paper. This is because at 157 nm illumination of old paper, compete bond

Alkiviadis C. Cefalas; Evangelia Sarantopoulou; Z. Kollia; Panagiotis Argitis

2001-01-01

216

The 2009 Port Wine Stain and Vascular Birthmarks Conference  

E-print Network

The 2009 Port Wine Stain and Vascular Birthmarks Conference was the Vascular Birthmarks Foundation. Nelson presented preliminary results of great interest to individuals with port wine stains. The current study results provide hope for a new combination of drug and laser therapy treatment that will remove

George, Steven C.

217

Selection and Application of Exterior Stains for Wood.  

National Technical Information Service (NTIS)

Exerior stains for wood protect the wood surface from sunlight and moisture. Because stains are formulated to penetrate the wood surface, they are not prone to crack or peel as can film forming finishes, such as paints. This publication describes the prop...

R. S. Williams, W. C. Feist

1999-01-01

218

Investigation of Methods for Removing Stains from Mortar and Concrete.  

National Technical Information Service (NTIS)

This investigation consisted of three parts: (a) a literature search to acquire information on the types of stains that can be expected and methods of removal; (b) evaluation of methods of removing stains from mortar specimens; and (c) evaluation of promi...

C. F. Derrington, R. L. Stowe, W. G. Miller

1968-01-01

219

Color of restorative materials after staining and bleaching.  

PubMed

This study determined the effect of a 10% carbamide peroxide bleaching agent on the removal of stain from restorative materials. Color changes (delta E*) of three restorative materials [compomer (Dyract); composite (TPH Spectrum); hybrid ionomer (Fuji II LC)] when exposed to juice/tea, chlorhexidine (CH), and water (control) for 120 hours were studied. Stained specimens were treated for two 2-hour periods with a bleaching agent (Platinum Tooth Whitening System) with and without the active ingredient. Color was measured at baseline, after staining, and after treatment using the CIE L*a*b* color system relative to CIE standard illuminant A (incandescent light) as measured by a reflection spectrophotometer. Means and standard deviations (n = 5) were calculated and data were analyzed by four-way ANOVA. All variables and interactions were statistically significant. Color changes caused by CH and water were not perceptible (delta E* < 3.3). After two 2-hour treatments, the following occurred with specimens stained with cranberry juice/tea: paste with and without active ingredient perceptibly changed color of stained composite. The stained hybrid ionomer perceptibly changed color after treatment with paste containing active ingredient but did not change after exposure to paste without active ingredient. The stained compomer was not perceptibly different with either treatment. Platinum successfully removed stains from the composite and hybrid ionomer tested. PMID:10823076

Fay, R M; Servos, T; Powers, J M

1999-01-01

220

Evaluation of fluorescent stains for visualizing extracellular DNA in biofilms.  

PubMed

Here we determine an optimal technique for the visualization of extracellular DNA in bacterial biofilms using the fluorescent eDNA stain TOTO-1 and the counterstain SYTO 60. This technique allows for more sensitive eDNA visualization than other fluorescent staining methods currently in use. PMID:25017901

Okshevsky, Mira; Meyer, Rikke Louise

2014-10-01

221

Development of an affordable dye-stained microalbuminuria screening test  

Microsoft Academic Search

Background. A simple spot test was developed, which al- lows quantification of microalbuminuria. Evaluation was carried out according to the ISO 15189 guidelines. Methods. Urine was spotted on cellulose acetate strips and stained using different sensitive protein binding dyes (ni- grosin, Coomassie Blue R-250, amido black). The colour intensity of the stained spots was quantified using a Kodak Image 450

Pierrot Lundimu Tugirimana; Joris R. Delanghe

2008-01-01

222

Alcian Blue Alizarin Red Skeletal Staining October 2003  

E-print Network

Alcian Blue ­ Alizarin Red Skeletal Staining October 2003 Eddy M. De Robertis 1. Dissect mice damage. 3. Replace 95% ethanol with Alcian blue staining solution for 1-3 days slowly rocking at room days. 4. Replace Alcian blue solution with 95% ethanol for 6 hours slowly rocking at room temperature

De Robertis, Eddy M.

223

Hydration of an Oriental Spruce (Picea orientalis) Pollen Grain  

NSDL National Science Digital Library

Time series during hydration of an oriental spruce (Picea orientalis) pollen grain. Rhodamine B stains the exine (red) while fluorescein diacetate crosses plasmalemmae and indicates esterase activity in the living cells (green). Swelling of the tube cell reduces the volume of the saccate air space and results in a loss of pollen buoyancy. Confocal extended depth of focus sections taken at (top to bottom) 1, 8, and 15 min after start of hydration.

C. John Runions (University of Victoria;Centre for Forest Biology ADR;POSTAL)

2004-03-09

224

Flow cytometric analysis of Eimeria tenella sporozoite populations exposed to salinomycin sodium in vitro: A comparative study using light and electron microscopy and an in vitro sporozoite invasion-inhibition test  

Microsoft Academic Search

Eimeria tenella sporozoites exposed to 100, 70, 60 and 50 µg salinomycin sodium (SAL)\\/ml medium 199 at 41° C and then stained with propidium iodide\\/fluorescein diacetate were analysed by means of flow cytometry (FCM). After 20 min exposure, they showed dose-dependent alterations in their size and shape, i.e. ballooning of most cells, and enhanced intracellular esterase activity as compared with

W. Raether; H. Mehlhorn; J. Hofmann; B. Briiu; K. Ehrlich

1991-01-01

225

Disinfection of seawater for hatchery aquaculture systems using electrolytic water treatment  

Microsoft Academic Search

A recently marketed electrolytic water treatment system (Hoshizaki) was evaluated for disinfection of seawater used in disease-prone high-intensity aquaculture systems. Bacterial plate counts (CFU), direct bacterial total counts using 4?,6? diamidino-2-phenylindole (DAPI) staining, and viable bacterial total counts using 6-carboxy fluorescein diacetate (6CFDA) showed complete inactivation of bacterial populations at an intensity of ?1.3 amp (?2.13 mg Cl l?1). This

Milko A. Jorquera; Gustavo Valencia; Mitsuru Eguchi; Masahiko Katayose; Carlos Riquelme

2002-01-01

226

Effect of docosahexaenoic acid on ras post-translational processing and localization in a transgenic mouse colonic cell line  

E-print Network

GAS CHROMOTOGRAPHY. . . . 77 APPENDIX 11 COMPLEXING FATTY ACIDS TO BSA. 81 APPENDIX 12 CELL NUMBER DETERMINATION . 84 APPENDIX 13 FLUORESCEIN DIACETATE - PROPIDIUM IODINE VIABILITY STAINING 85 APPENDIX 14 COLUMN CHROMATOGRAPHY TO REMOVE NEUTRAL..., subconfluent cells were treated with 50 ItM DHA or LA complexed to fatty acid free BSA (Holub et al. 1971) for 72 h (Appendix 11). Cells treated with media only were used as a control for cell number, viability, and fatty acid incorporation. Following...

Collett, Esther Dick

2012-06-07

227

Change in chain stiffness in viscometric and ultracentrifugal fields: Cellulose diacetate in N, N-dimethylacetamide dilute solution  

NASA Astrophysics Data System (ADS)

The molecular characteristics and the chain stiffness were investigated for fractionated samples of cellulose diacetate (CDA, degree of substitution DS = 2.40) in N, N-dimethylacetamide (DMAc) through the partial specific volume, the viscometric, the sedimentation velocity, and the sedimentation equilibrium measurements at 30 °C. It was found that CDA dispersed molecularly in DMAc under the external field such as in the viscometric and the sedimentation experiments. The molecular weight dependence of the intrinsic viscosity [?] and the sedimentation coefficient at infinite dilution s0 of the single CDA molecule were expressed by the relation [?] = 1.10×10-2Mw0.85 (cm3 g-1) and s0=2.25×10-14Mw0.20 (s), which exhibit the stiff, or semiflexible chain nature. The semiflexible chain parameters were evaluated by the Yamakawa-Fujii (YF) theory of the unperturbed wormlike cylinder model, first via a combination of the viscosity and the partial specific volume data (method A) and second via a combination of the sedimentation and the partial specific volume data (method B). Method A gave the chain parameters that q = 8.0 nm, ML = 523 nm-1, and d = 0.89 nm, whereas method B gave q = 48 nm, ML = 560 nm-1, and d = 0.93 nm. Here q is the persistence length, ML is the molecular weight per unit contour length, and d is the chain diameter of the wormlike cylinder model. Methods A and B deduce, independent of the method, the definite ML and d values, which are very consistent with the ordinary reported values. However, q estimated by the two methods differs by about six times the other. This fact suggests that the CDA molecule in the ultracentrifugal field has a conformation different from that in the viscometric shear field: The CDA chain may be highly stiff in the ultracentrifuge because of the situation that the adjacent glucose residues are stuck in a rigid conformation by the double stapled hydrogen bonds between the intramolecular hydroxyls and oxygens.

Kawanishi, Hiroyuki; Tsunashima, Yoshisuke; Okada, Shinichi; Horii, Fumitaka

1998-04-01

228

Novel Process for Laser Stain Removal from Archaeological Oil Paintings  

NASA Astrophysics Data System (ADS)

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.

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

2013-03-01

229

Automated detection of cells from immunohistochemically-stained tissues: application to Ki-67 nuclei staining  

NASA Astrophysics Data System (ADS)

An automated cell nuclei detection algorithm is described to be used for the quantification of immunohistochemicallystained tissues. Detection and segmentation of positively stained cells and their separation from the background and negatively-stained cells is crucial for fast, accurate, consistent and objective analysis of pathology images. One of the major challenges is the identification, hence accurate counting of individual cells, when these cells form clusters. To identify individual cell nuclei within clusters, we propose a new cell nuclei detection method based on the well-known watershed segmentation, which can lead to under- or over-segmentation for this problem. Our algorithm handles oversegmentation by combining H-minima transformed watershed algorithm with a novel region merging technique. To handle under-segmentation problem, we develop a Laplacian-of-Gaussian (LoG) filtering based blob detection algorithm, which estimates the range of the scales from the image adaptively. An SVM classifier was trained in order to separate non-touching single cells and touching cell clusters with five features representing connected region properties such as eccentricity, area, perimeter, convex area and perimeter-to-area ratio. Classified touching cell clusters are segmented with the H-minima based watershed algorithm. The resulting over-segmented regions are improved with the merging algorithm. The remaining under-segmented cell clusters are convolved with LoG filters to detect the cells within them. Cell-by-cell nucleus detection performance is evaluated by comparing computer detections with cell locations manually marked by eight pathology residents. The sensitivity is 89% when the cells are marked as positive at least by one resident and it increases to 99% when the evaluated cells are marked by all eight residents. In comparison, the average reader sensitivity varies between 70% +/- 18% and 95% +/- 11%.

Cinar Akakin, Hatice; Kong, Hui; Elkins, Camille; Hemminger, Jessica; Miller, Barrie; Ming, Jin; Plocharczyk, Elizabeth; Roth, Rachel; Weinberg, Mitchell; Ziegler, Rebecca; Lozanski, Gerard; Gurcan, Metin N.

2012-03-01

230

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

PubMed Central

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

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

2013-01-01

231

Dietary staining in vitro by mouthrinses as a comparative measure of antiseptic activity and predictor of staining in vivo.  

PubMed

Extrinsic staining of teeth is a side-effect of some antiseptic mouthrinses. However, few of the many rinse products available to the general public have been investigated for their propensity to cause staining. Dietary factors play an aetiological role in staining and have been used in vitro to study and compare the activity of rinses. The aim of this study was to assess rinse products for staining in vitro and, through the staining reaction, to compare the activity of products containing the same ingredients. Perspex blocks, with or without saliva pretreatment, were soaked in rinses for 2 min, washed and placed in a standard tea solution for 60 min and then the optical density (OD) read on a spectrophotometer. The cycle was repeated 10 times for saliva and 17 times for no saliva specimens or until the maximum OD was exceeded. A series of three separate experiments was performed by this method. The maximum OD was not exceeded by any product before seven passages and therefore data were compared at six passages. For most products OD increased with saliva pretreatment. Some cetylpyridinium chloride (CPC) rinses stained comparably to a chlorhexidine rinse. CPC rinses, most of which contained the same concentration of the antiseptic, varied considerably in their propensity to induce staining and one was little different to water controls. A 0.1% chlorhexidine rinse stained slightly more than a 0.2%. A phenolic/essential oil product produced some staining but zinc, triclosan and other essential oil rinses did not stain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7738271

Addy, M; Mahdavi, S A; Loyn, T

1995-04-01

232

Growth Inhibitory Effect of (E)-2,4-bis(p-hydroxyphenyl)-2-Butenal Diacetate through Induction of Apoptotic Cell Death by Increasing DR3 Expression in Human Lung Cancer Cells  

PubMed Central

The Maillard Reaction Products (MRPs) are chemical compounds which have been known to be effective in chemoprevention. Death receptors (DR) play a central role in directing apoptosis in several cancer cells. In our previous study, we demonstrated that (E)-2,4-bis(p-hydroxyphenyl)-2-butenal, a MRP product, inhibited human colon cancer cell growth by inducing apoptosis via nuclear factor-?B (NF-?B) inactivation and G2/M phase cell cycle arrest. In this study, (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate, a new (E)-2,4-bis(p-hydroxyphenyl)-2-butenal derivative, was synthesized to improve their solubility and stability in water and then evaluated against NCI-H460 and A549 human lung cancer cells. (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate reduced the viability in both cell lines in a time and dose-dependent manner. We also found that (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate increased apoptotic cell death through the upregulation of the expression of death receptor (DR)-3 and DR6 in both lung cancer cell lines. In addition to this, the transfection of DR3 siRNA diminished the growth inhibitory and apoptosis inducing effect of (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate on lung cancer cells, however these effects of (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate was not changed by DR6 siRNA. These results indicated that (E)-2,4-bis(p-hydroxyphenyl)-2-butenal diacetate inhibits human lung cancer cell growth via increasing apoptotic cell death by upregulation of the expression of DR3. PMID:24009847

Lee, Ung-Soo; Ban, Jung Ok; Yeon, Eung Tae; Lee, Hee Pom; Udumula, Venkatareddy; Ham, Young Wan; Hong, Jin Tae

2012-01-01

233

Western Blot of Stained Proteins from Dried Polyacrylamide Gels  

NASA Technical Reports Server (NTRS)

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.

Gruber, Claudia; Stan-Lotter, Helga

1996-01-01

234

Preoperative methylene blue staining of galactographically suspicious breast lesions.  

PubMed

Microdochectomy is the standard treatment of galactographically suspicious breast lesions. Precise preoperative marking of the suspicious duct and intraductal lesions facilitates selective minimal-volume microdochectomy. Methylene blue dye staining fulfills this criterion. A retrospective review of our experience of preoperative methylene blue staining in 30 patients with unilateral spontaneous nonlactiferous single duct nipple discharge operated on during 1986-1995 in the Oulu University Hospital for galactographically suspicious breast lesions. Galactography was successful in 29 out of 30 (93.3%) cases. Preoperative methylene blue staining was attempted in all cases on the day of surgery and it was successful in 22 (73.3%) cases making subsequent selective minimal-volume microdochectomy easy to perform. The failure of methylene blue staining led to quadrantectomy in 4 cases and smaller breast resections in the remaining 4 cases. Preoperative methylene blue dye staining crucially facilitates selective minimal-volume microdochectomy. An interval between primary galactography and later methylene blue staining leads to failures in approximately one quarter of the cases. A higher success rate would necessitate scheduling the microdochectomy on the same day as the primary galactography (and the subsequent methylene blue staining in suspicious cases). PMID:9412841

Saarela, A O; Kiviniemi, H O; Rissanen, T J

1997-01-01

235

Staining with methylthioninium chloride for the diagnosis of fungal keratitis  

PubMed Central

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

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

2013-01-01

236

Dual role of acidic diacetate sophorolipid as biostabilizer for ZnO nanoparticle synthesis and biofunctionalizing agent against Salmonella enterica and Candida albicans.  

PubMed

In the present study, a yeast species isolated from CETP, Vellore, Tamilnadu was identified as Cryptococcus sp. VITGBN2 based on molecular techniques and was found to be a potent producer of acidic diacetate sophorolipid in mineral salt media containing vegetable oil as additional carbon source. The chemical structure of the purified biosurfactant was identified as acidic diacetate sophorolipid through GC-MS analysis. This sophorolipid was used as a stabilizer for synthesis of zinc oxide nanoparticles (ZON). The formation of biofunctionalized ZON was characterized using UV-visible spectroscopy, XRD, scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy. The antimicrobial activities of naked ZON and sophorolipid functionalized ZON were tested based on the diameter of inhibition zone in agar well diffusion assay, microbial growth rate determination, protein leakage analysis, and lactate dehydrogenase assay. Bacterial pathogen Salmonella enterica and fungal pathogen Candida albicans showed more sensitivity to sophorolipid biofunctionalized ZON compared with naked ZON. Among the two pathogens, S. enterica showed higher sensitivity towards sophorolipid biofunctionalized ZON. SEM analysis showed that cell damage occurred through cell elongation in the case of S. enterica, whereas cell rupture was found to occur predominantly in the case of C. albicans. This is the first report on the dual role of yeastmediated sophorolipid used as a biostabilizer for ZON synthesis as well as a novel functionalizing agent showing antimicrobial property. PMID:24150496

Basak, Geetanjali; Das, Devlina; Das, Nilanjana

2014-01-01

237

[Changes of various proteins in older blood stains].  

PubMed

26 unequally old groups of blood stains up to 180 days old were examined for the presence of albumin, Ig A, Ig G immunoglobulins, beta 1 C globulin and transferrin. To ensure different external conditions, each group was composed so as to contain blood stains of the same age but preserved under different conditions and on different types of underlying material. Albumin and Ig G immunoglobulin proved to be among the most stable ones. However, due to variations in the results the practical uses of this type of blood stain age estimation appear to be problematic. PMID:88276

Klír, P

1979-05-01

238

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

PubMed Central

Abstract. 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. PMID:22734736

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

2012-01-01

239

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

SciTech Connect

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.

Colotelo, Alison HA; Cooke, Steven J.

2011-05-01

240

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

241

7 CFR 28.442 - Middling Yellow Stained Color.  

Code of Federal Regulations, 2010 CFR

7 ? Agriculture ? 2 ? 2010-01-01 ? 2010-01-01 ? false ? Middling Yellow Stained Color. ? 28.442 ? Section 28.442 ? Agriculture ? Regulations of the Department of Agriculture ? AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE ?...

2010-01-01

242

Immunohistochemical staining for ranaviruses Introduction: Ranaviruses negatively impact amphibian populations  

E-print Network

(Trachemys scripta elegans) that were challenged with 4 different FV3-like ranavirus isolates (FV3, isolate, no staining was observed in the tissues of the red eared slider (Trachemys scripta elegans; Fig. 3

Gray, Matthew

243

Removing foxing stains from old paper at 157 nm  

Microsoft Academic Search

Using a molecular fluorine laser at 157nm foxing stains were removed successfully from a 16th century old paper. Laser cleaning of stains and foxing from old paper manuscripts is far more effective at 157nm in comparison to different wavelengths without leaving any yellowish after-effect on the paper. This is because at 157nm illumination of old paper, complete bond breaking of

E. Sarantopoulou; Z. Samardzija; S. Kobe; Z. Kollia; A. C. Cefalas

2003-01-01

244

Multispectral image enhancement for H&E stained pathological tissue specimens  

Microsoft Academic Search

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

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

2008-01-01

245

Accumulation of supramolecular nanoparticles self-assembled from a bola-shaped cytidylic acid-appended fluorescein dye in cell nuclei.  

PubMed

We examined the cellular uptake of the nanoparticles self-assembled from a bola-shaped cytidylic acid-appended fluorescein derivative (C-FLU-C). The accumulation of fluorescence in the Caco-2 cell nucleus was observed mainly after the plateau phase of cell growth, indicating that C-FLU-C permeated the nuclear envelope without nuclear-localizing tags. PMID:25000245

Iwaura, Rika; Shirai, Mutsumi; Yoshida, Kaname; Ohnishi-Kameyama, Mayumi

2014-08-25

246

Quantitative measurement of retinal blood flow in human beings by application of digital image-processing methods to television fluorescein angiograms  

Microsoft Academic Search

A method is presented that allows the quantitative determination of the blood flow in retinal arteries in human beings. Television fluorescein angiograms are used as input. This method does not need any gauge procedures since all the necessary information is taken from the image itself. Also, the patients' eye movements do not introduce errors because their influence is removed by

P. R. Preußner; G. Richard; O. Darrelmann; J. Weber; I. Kreissig

1983-01-01

247

Behaviour of the iris vasculature in central retinal vein occlusion: a fluorescein angiographic study of the vascular response of the retina and the iris  

Microsoft Academic Search

The findings in iris fluorescein angiograms of 48 eyes with central retinal vein occlusion (CRVO) were correlated with the predominant retinal vascular response. In 24 eyes with the non-ischaemic type of CRVO there were no or only minimal iris vessel changes, whereas in all 24 eyes with ischaemic type of CRVO there was iris vessel dilatation and leakage with or

L Laatikainen; R K Blach

1977-01-01

248

Comparing modified papanicolaou stain with ayoub-shklar and haematoxylin-eosin stain for demonstration of keratin in paraffin embedded tissue sections  

PubMed Central

Aim: The aim of the present study was to stain the known keratin containing tissues by Haematoxylin and eosin stain (H-E), ayoub-shklar (A/S) and modified Papanicolaou (PAP) stain and to compare the efficacy of modified PAP staining procedure with that of A/S stain and H-E staining technique, so as to device a staining procedure which is easy and effective for keratin. Materials and Methods: A Total Number of 60 paraffin embedded tissue sections of known keratin containing tissues including normal keratinized oral mucosa (NKOM), Keratinized Odontogenic Keratocyst (OKC), Verrucous Carcinoma (VC) and Well differentiated Squamous Cell Carcinoma (WDSCC) were taken and 3 sections of 4 microns thickness of each block were cut and stained with above mentioned three stains. Results: Surface keratin was stained distinctly and uniformly in all the three staining techniques in NKOM, OKC, and VC and WDSCC. But results were statistically significant in WDSCC when Amount of keratin pearls and Pattern of staining were compared in all 3 staining procedures and p value was p=0.000 and p=0.001 respectively. Conclusion: Based on the above findings we conclude that the efficacy of modified PAP is comparable with that of H-E stain and A-S stain for surface keratin and thus be used effectively to stain Surface keratin. Whereas to know the exact pattern of cytokeratin expression in SCC a more sensitive tool like immunohistochemical method can be applied. PMID:23798825

Ramulu, Surekha; Kale, Alka D; Hallikerimath, Seema; Kotrashetti, Vijayalaxmi

2013-01-01

249

Reliability of a rapid hematology stain for sputum cytology*  

PubMed Central

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

Goncalves, Jessica; Pizzichini, Emilio; Pizzichini, Marcia Margaret Menezes; Steidle, Leila John Marques; Rocha, Cristiane Cinara; Ferreira, Samira Cardoso; Zimmermann, Celia Tania

2014-01-01

250

The stain prevention efficacy of two tooth whitening dentifrices.  

PubMed

An 8-week randomized, double-blind, parallel group clinical study was conducted to assess the extrinsic stain prevention efficacy of three commercially available dentifrices: 1) a dentifrice containing 0.243% sodium fluoride with copolymer, tetrasodium pyrophosphate, and sodium tripolyphosphate in a silica base (Product 1); 2) a dentifrice containing 0.243% sodium fluoride, baking soda and peroxide, tetrasodium pyrophosphate, and sodium tripolyphosphate in a silica base (Product 2); and 3) a dentifrice containing 0.243% sodium fluoride in a silica base (Product 3). After the collection of baseline stain scores by a trained examiner and a subsequent oral prophylaxis, 126 volunteers were randomized to one of the three treatment groups (balanced for composite extrinsic stain scores). Throughout the 8-week treatment period, subjects brushed their teeth twice daily with their assigned dentifrice. At baseline, 4-, and 8-week evaluations, extrinsic dental stain was measured on the facial surfaces of the six maxillary anterior teeth and on the facial and lingual surfaces of the six mandibular anterior teeth using the Lobene Index. A total of 120 subjects completed the study. No adverse events were reported, and subjects who discontinued the study did so for reasons unrelated to the dentifrices. At the 4-week evaluation, composite stain scores were statistically significantly lower (P < .05) for both Product 1 (44.9%) and for Product 2 (34.6%) relative to Product 3. At the 8-week evaluation, composite stain scores were statistically significantly lower (P < .05) for both Product 1 (28.4%) and for Product 2 (29.6%) relative to Product 3. The results of this clinical study demonstrate that both dentifrices, one containing 0.234% sodium fluoride with copolymer, tetrasodium pyrophosphate, and sodium tripolyphosphate in a silica base; and one with 0.243% sodium fluoride with copolymer, tetrasodium pyrophosphate, and sodium tripolyphosphate in a silica base are more effective in preventing natural extrinsic stain formation on teeth as compared with a marketed control dentifrice. PMID:12244740

Ayad, Farid; De Sciscio, Peter; Stewart, Bernal; De Vizio, William; Petrone, Margaret E; Volpe, Anthony R

2002-08-01

251

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

E-print Network

protein gel stain, Coomassie brilliant blue or silver stain. We typically use .igure 1. Staining 365 stain exhibits a bright blue fluo- rescence when excited with UV light, (e.g., with a standard UV. Materials Required but Not Provided Polystyrene staining dish Ethanol, spectroscopy grade Glacial acetic

Lebendiker, Mario

252

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

E-print Network

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

Mecham, Robert

253

Correlation between Octopus perimetry and fluorescein angiography after strontium-90 plaque brachytherapy for subfoveal exudative age related macular degeneration  

PubMed Central

AIM—To evaluate the correlation between the central visual field and changes in fluorescein angiography and fundus photography in patients treated with strontium plaque radiotherapy for subfoveal exudative age related macular degeneration (AMD).?METHODS—Octopus program 34 automated static perimetry, fluorescein angiography, and colour fundus photography were performed on 19 patients at baseline and at 12 months after strontium-90 plaque therapy. A schematic picture outlining the areas of hyperfluorescent neovascular membranes and subretinal blood was drawn of a projected 30° fundus fluorescein angiogram. This drawing was superimposed on the size adjusted Octopus visual field. The changes in retinal sensitivity were calculated and related to angiographic changes.?RESULTS—Three of the 19 patients had a reliability factor (RF) >15% and were excluded from further analysis. In the remaining 16 patients the mean defect (MD) and loss variance (LV) values remained unchanged in patients showing regression of the choroidal neovascular membrane (CNVM) to irradiation at 12 months. MD was 7.7 (SD 1.7) at baseline and 7.6 (1.9) at 12 months (p = 0.86), and LV was 32.6 (13.9) at baseline and 32.4 (15.7) at 12 months (p = 0.94). However, in patients with progression of the CNVM at 12 months, both the MD and LV increased significantly during the 12 month follow up (MD from 7.3 (2.9) to 13.1 (3.6) ( p = 0.05) and LV from 31.0 (22.9) to 71.8 (24.1) (p = 0.017)). When comparing the mean retinal sensitivity in the area of the primary CNVM (including classic, occult, and haemorrhagic components), the results were analogous: in patients with a regression of the CNVM after irradiation the mean sensitivity remained almost unchanged. It was 10.3 (6.4) dB at baseline and 9.4 (7.3) dB at 12 months (p = 0.58). In five out of 11 patients (45%) with regression of the CNVM, the mean retinal sensitivity even improved by 2.0-5.0 dB in the area of the original lesion during follow up. Instead, in patients showing progression of the CNVM at 12 months, there was a significant loss in mean retinal sensitivity—from 9.9 (4.6) dB at baseline to 1.0 (1.1) dB at 12 months (p = 0.019). The mean retinal sensitivity in the area of the irradiated but clinically normal retina during follow up was not significantly altered (21.5 dB at baseline, 19.7 dB at 12 months (p = 0.10)).?CONCLUSIONS—Regression of subfoveal choroidal membranes in AMD after focal strontium irradiation is connected with stabilisation or even improvement of retinal sensitivity in central visual field measured by automated perimetry. Strontium plaque irradiation does not change the sensitivity in clinically normal paramacular retina during a 12 month follow up.?? Keywords: choroidal neovascular membranes; age related macular degeneration; Octopus perimetry; strontium PMID:9924368

Jaakkola, A.; Vesti, E.; Immonen, I.

1998-01-01

254

Application of a Vital Fluorescent Staining Method for Simultaneous, Near-Real-Time Concentration Monitoring of Two Bacterial Strains in an Atlantic Coastal Plain Aquifer in Oyster, Virginia  

PubMed Central

Two differentially labeled bacterial strains were monitored in near-real time during two field-scale bacterial transport experiments in a shallow aquifer in July 2000 and July 2001. Comamonas sp. strain DA001 and Acidovorax sp. strain OY-107 were grown and labeled with the vital fluorescent stain TAMRA/SE (5 [and -6]-carboxytetramethylrhodamine, succinimidyl ester) or CFDA/SE (5 [and -6]-carboxyfluorescein diacetate, succinimidyl ester). Fluorescently labeled cells and a conservative bromide tracer were introduced into a suboxic superficial aquifer, followed by groundwater collection from down-gradient multilevel samplers. Cells were enumerated in the field by microplate spectrofluorometry, with confirmatory analyses for selected samples done in the laboratory by epifluorescence microscopy, flow cytometry, and ferrographic capture. There was general agreement in the results from all of the vital-stain-based enumeration methods, with differences ranging from <10% up to 40% for the analysis of identical samples between different tracking methods. Field analysis by microplate spectrofluorometry was robust and efficient, allowing thousands of samples to be analyzed in quadruplicate for both of the injected strains. The near-real-time data acquisition allowed adjustments to the predetermined sampling schedule to be made. The microplate spectrofluorometry data sets for the July 2000 and July 2001 experiments allowed the transport of the injected cells to be related to the site hydrogeology and injection conditions and enabled the assessment of differences in the transport of the two strains. This near-real-time method should prove effective for a number of microbial ecology applications. PMID:15006793

Fuller, Mark E.; Mailloux, Brian J.; Streger, Sheryl H.; Hall, James A.; Zhang, Pengfei; Kovacik, William P.; Vainberg, Simon; Johnson, William P.; Onstott, Tullis C.; DeFlaun, Mary F.

2004-01-01

255

Methods of biological dosimetry employing chromosome-specific staining  

DOEpatents

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.

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

2000-01-01

256

Methods And Compositions For Chromosome-Specific Staining  

DOEpatents

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.

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

2003-08-19

257

Centromere and cytoplasmic staining pattern recognition: a local approach.  

PubMed

Autoimmune diseases are very serious and also invalidating illnesses. The benchmark procedure for their diagnosis is the indirect immunofluorescence (IIF) assay performed on the HEp-2 substrate. Medical doctors first determine the fluorescence intensity exhibited by HEp-2 wells and then report the staining pattern. Despite its pivotal role, IIF is affected by inter- and intra-laboratory variabilities demanding for the development of computer-aided-diagnosis tools supporting medical doctor decisions. With reference to staining pattern recognition, state-of-the-art approaches recognize five main patterns characterized by well-defined cell edges. These approaches are based on cell segmentation, a task that recent work suggests to be harder than the classification itself. In this paper, we extend the panel of detectable HEp-2 staining patterns, introducing the recognition of centromere and cytoplasmic patterns, which have a high specific match with certain autoimmune diseases, from other stainings. Since image segmentation algorithms fail on these samples, we developed a classification system integrating local descriptors and the bag of visual word approach, which represents image contents without the burden of segmentation. We tested our approach on a large dataset of HEp-2 images with high variability in both fluorescence intensity and staining patterns correctly recognizing the 97.12 % of samples. The system has also been validated in a daily routine fashion on 108 consecutive IIF analyses of hospital outpatients and inpatients, achieving an accuracy rate of 97.22 %. PMID:23877232

Iannello, Giulio; Onofri, Leonardo; Soda, Paolo

2013-12-01

258

Histochemical staining of Arabidopsis thaliana secondary cell wall elements.  

PubMed

Arabidopsis thaliana is a model organism commonly used to understand and manipulate various cellular processes in plants, and it has been used extensively in the study of secondary cell wall formation. Secondary cell wall deposition occurs after the primary cell wall is laid down, a process carried out exclusively by specialized cells such as those forming vessel and fiber tissues. Most secondary cell walls are composed of cellulose (40-50%), hemicellulose (25-30%), and lignin (20-30%). Several mutations affecting secondary cell wall biosynthesis have been isolated, and the corresponding mutants may or may not exhibit obvious biochemical composition changes or visual phenotypes since these mutations could be masked by compensatory responses. Staining procedures have historically been used to show differences on a cellular basis. These methods are exclusively visual means of analysis; nevertheless their role in rapid and critical analysis is of great importance. Congo red and calcofluor white are stains used to detect polysaccharides, whereas Mäule and phloroglucinol are commonly used to determine differences in lignin, and toluidine blue O is used to differentially stain polysaccharides and lignin. The seemingly simple techniques of sectioning, staining, and imaging can be a challenge for beginners. Starting with sample preparation using the A. thaliana model, this study details the protocols of a variety of staining methodologies that can be easily implemented for observation of cell and tissue organization in secondary cell walls of plants. PMID:24894795

Pradhan Mitra, Prajakta; Loqué, Dominique

2014-01-01

259

Amyloid Histology Stain for Rapid Bacterial Endospore Imaging ? †  

PubMed Central

Bacterial endospores are some of the most resilient forms of life known to us, with their persistent survival capability resulting from a complex and effective structural organization. The outer membrane of endospores is surrounded by the densely packed endospore coat and exosporium, containing amyloid or amyloid-like proteins. In fact, it is the impenetrable composition of the endospore coat and the exosporium that makes staining methodologies for endospore detection complex and challenging. Therefore, a plausible strategy for facile and expedient staining would be to target components of the protective surface layers of the endospores. Instead of targeting endogenous markers encapsulated in the spores, here we demonstrated staining of these dormant life entities that targets the amyloid domains, i.e., the very surface components that make the coats of these species impenetrable. Using an amyloid staining dye, thioflavin T (ThT), we examined this strategy. A short incubation of bacillus endospore suspensions with ThT, under ambient conditions, resulted in (i) an enhancement of the fluorescence of ThT and (ii) the accumulation of ThT in the endospores, affording fluorescence images with excellent contrast ratios. Fluorescence images revealed that ThT tends to accumulate in the surface regions of the endospores. The observed fluorescence enhancement and dye accumulation, coupled with the sensitivity of emission techniques, provide an effective and rapid means of staining endospores without the inconvenience of pre- or posttreatment of samples. PMID:21653779

Xia, Bing; Upadhyayula, Srigokul; Nunez, Vicente; Landsman, Pavel; Lam, Samuel; Malik, Harbani; Gupta, Sharad; Sarshar, Mohammad; Hu, Jingqiu; Anvari, Bahman; Jones, Guilford; Vullev, Valentine I.

2011-01-01

260

Dissociation kinetics of 1,10-diaza-4,7,13,16-tetraoxacyclooctadecane-N,N'-diacetic acid complexes of lanthanides  

SciTech Connect

The dissociation kinetics of 1,10-diaza-4,7,13,16-tetraoxacyclooctadecane-N,N'-diacetic acid (K22DA) complexes of lanthanide(III) ions were studied in perchloric acid and other media, over the concentration range 5 x 10 U to 7.5 x 10 T M and at a constant ionic strength of 0.1 M (LiClO4). Copper (II) was used as the scavenger of free ligand, and the rates of dissociation of these complexes have been found to be independent of (CuS ). All the complexes exhibit acid-dependent and acid-independent pathways in a manner similar to those of LnCyDTA complexes (CyDTA = trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetate ion). Unlike lanthanide (Ln) complexes of 1,7-diaza-4,10,13-trioxacyclopentadecane-N,N'-diacetic acid (K21DA), LnK22DA complex dissociation rates are independent of (anion) and (electrolyte). There is also no general-acid catalysis. The overall results are compared with those of complexes that are structural analogues such as LnK21Da , LnMEDTA (MEDTA = N-methyl-ethylenediamine-N,N,N'-triacetate ion), and LnCyDTA . A rationalization is given to account for similarities and differences with respect to observed kinetic characteristics. A number of postulates concerning the relationship between thermodynamic and kinetic stabilities and the detailed reaction mechanisms of lanthanide complxes of polyamino polyacetate ligands are proposed. 19 references, 1 figure, 7 tables.

Chang, C.A.; Sekhar, V.C.

1987-06-17

261

Optical waveguide fabricated by ion-exchange using stain method  

NASA Astrophysics Data System (ADS)

Optical waveguides were prepared by the incorporation of silver or copper ions using the classical staining. We used commercially available soda-lime silicate and borosilicate glasses as substrates. Silver or copper stain was applied on a side of the glass substrates. The substrates were heat-treated at elevated temperature for various times. The treated glasses were optically clear and almost colorless except for a few samples stained for longer time. This indicates that silver and copper metal nanoparticles and Cu2O nanoparticles causing coloration of glasses were not formed in the glass substrates. The ion-incorporation process was approximately controlled by the diffusion of ions. We observed the propagation of 633 nm laser radiation by a prism coupling method showing that the glass surface region plays a role of waveguide. Refractive index change more than 0.01 at 633 nm was achieved in the waveguide layers.

Kadono, Kohei; Suetsugu, Tatsuya; Ohtani, Takeshi; Kominami, Norimasa; Takada, Minoru; Einishi, Toshihiko; Tarumi, Takashi

2005-04-01

262

Colorimetry for the stain technologist. I. The specification of color.  

PubMed

This paper describes color specification for the stain technologist. The principles of color stimulus specification are reviewed in terms of the conventions of the Commission Internationale de l'Eclairage (CIE). The text is largely self-contained and has been written so that it can be understood easily by a reader with no prior knowledge of color science. The paper starts with definitions of color and related psychological, psychophysical and colorimetric terms. X, Y, Z color space is described. It is shown that any color stimulus may be unambiguously defined in terms of a set of three numbers. The CIE 1931 Chromaticity Diagram is described. Worked examples are given for the calculation of tristimulus values and chromaticity coordinates using three different illuminants. The usefulness of color specification is illustrated by a number of examples using Romanowsky stained blood cells or Papanicolaou stained epithelial cells from the uterine cervix. PMID:6208644

Marshall, P N; Galbraith, W

1984-07-01

263

Peroxisomes: 40 years of histochemical staining, personal reminiscences.  

PubMed

The historical circumstances that led to the discovery of the 3,3'-diamino-benzidine (DAB) method for staining of peroxisomes 40 years ago are reviewed. In the course of studies on the uptake and absorption of horse radish peroxidase in mammalian liver, in sections incubated for detection of peroxidase activity in DAB, it was noted that peroxisomes also stained positively for peroxidase activity. Subsequently, it was revealed that the peroxidatic activity of catalase, which is abundantly present in peroxisomes, is responsible for that staining. This notion was confirmed in quantitative biochemical studies with crystalline beef liver catalase and in tracer studies using catalase as an ultrastructural tracer. The application of the DAB method led to the discovery of peroxisomes as a ubiquitous eukaryotic cell organelle, attracting great interest in their investigation in biomedical research. PMID:19219449

Dariush Fahimi, H

2009-04-01

264

Morphofunctional analysis of human platelets by vital staining.  

PubMed

We developed a method for differential staining of human platelets preserving their functional activity based on vital fl uorochrome stains trypafl avin and acridine orange. Platelets stained with trypafl avin and acridine orange exhibited under a fl uorescent microscope green fl uorescence of the cytoplasm and red-orange fl uorescence of the granules. Morphofunctional analysis of platelets was carried out on the cells from donor blood, donor concentrated platelets, and cells from hematological patients and patients with thromboembolic complications. Populations with low (16 %) and high (2 %) morphofunctional activities of platelets were detected among donors. The morphofunctional parameters of platelets were sharply reduced in hematological patients with the hemorrhagic syndrome and elevated signifi cantly in patients with thromboembolic complications in comparison with donors. The method seemed to be effective for evaluating the platelet quality in donor and patients' blood components. PMID:24771387

Makarov, M S; Kobzeva, E N; Vysochin, I V; Borovkova, N V; Khvatov, V T

2014-01-01

265

A staining protocol for identifying secondary compounds in Myrtaceae1  

PubMed Central

• 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

Retamales, Hernan A.; Scharaschkin, Tanya

2014-01-01

266

Feasibility of digitally stained multimodal confocal mosaics to simulate histopathology  

NASA Astrophysics Data System (ADS)

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.

Gareau, Daniel S.

2009-05-01

267

Feasibility of digitally stained multimodal confocal mosaics to simulate histopathology.  

PubMed

Fluorescence confocal mosaicing microscopy of tissue biopsies stained with acridine orange has been shown to accurately identify tumors and with an overall sensitivity of 96.6% and specificity of 89.2%. However, fluorescence shows only nuclear detail similar to hematoxylin in histopathology and does not show collagen or cytoplasm, which may provide necessary negative contrast information similar to eosin used in histopathology. Reflectance mode contrast is sensitive to collagen and cytoplasm without staining. To further improve sensitivity and specificity, digitally stained confocal mosaics combine confocal fluorescence and reflectance images in a multimodal pseudo-color image to mimic the appearance of histopathology with hematoxylin and eosin and facilitate the introduction of confocal microscopy into the clinical realm. PMID:19566342

Gareau, Daniel S

2009-01-01

268

Chromosome-specific staining to detect genetic rearrangements  

DOEpatents

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.

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

2013-04-09

269

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

E-print Network

, such as SYPRO Ruby protein gel stain, Coomassie brilliant blue or silver stain. We typically use disposable for at least 6 months. Materials Required but Not Provided Polystyrene staining dish Ethanol, spectroscopy

Lebendiker, Mario

270

Toward Digital Staining using Imaging Mass Spectrometry and Random Forests  

PubMed Central

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

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

2009-01-01

271

Chemical aspects of santalin as a histological stain.  

PubMed

Recent research on the chemical nature of the red dyes isolated from Pterocarpus santalinus and certain West African plants, viz., Baphia nitida, Pterocarpus osun and Pterocarpus soyauxii, have been reviewed. P. santalinus contains santalins A, B and C, but no santarubin. Santalins and santarubins have been found in P. osun, P. soyauxii and B. nitida. The structural formulae of the santalins are presented and their differences from santarubins indicated. Santalins A and B have some similarities in structure with hematein. This is probably responsible for their staining properties; the possible mechanism of staining is discussed. PMID:6166100

Banerjee, A; Mukherjee, A K

1981-03-01

272

Novel drug delivery system of hollow mesoporous silica nanocapsules with thin shells: preparation and fluorescein isothiocyanate (FITC) release kinetics.  

PubMed

Core-shell nanoparticles of Au@silica with a diameter of approximate 45-60 nm and wall thickness in range of 3-10 nm were synthesized by using 40 and 50 nm gold nanoparticles as the templates. The mesoporous particles are regulated by 3-aminopropyltrimethoxysilane addition. Hollow mesoporous silica nanocapsules (HMSNs) were prepared by using sodium cyanide to dissolve the gold cores. The characterization of Au@silica and HMSNs by transmission electronic microscope indicated that the silica shells were uniform and smooth, and also the porosity was proved by fluorescein isothiocyanate (FITC) release experiments. The ratio of hollow core to HMSNs is more than 70%. HMSNs were subsequently used as drug carrier to investigate FITC (as a model drug) release behaviors in vitro. Fluorescent spectrometry was performed to determine the release kinetics from the HMSNs. The release profiles are significantly different as compared with the control (free FITC), which show that HMSNs are good drug carriers to control drug release, and have high potential in therapeutic drugs delivery in future applications. PMID:17420116

Liu, Yiyao; Miyoshi, Hirokazu; Nakamura, Michihiro

2007-08-01

273

Renal cortical basolateral Na+/HCO3- cotransporter: II. Detection of conformational changes with fluorescein isothiocyanate labeling.  

PubMed

Fluorescein isothiocyanate (FITC) fluorescently labels amino groups and has been useful in detecting conformational changes in transport proteins through quenching or enhancement of the fluorescence signal upon exposure of protein to substrates. Solubilized renal basolateral membrane proteins, enriched in Na+/HCO3- cotransporter activity, were reconstituted into liposomes and treated with FITC or its nonfluorescent analogue PITC (phenyl isothiocyanate). In the absence of Na+ and HCO3-, incubation of proteoliposomes with PITC or FITC significantly inhibited cotransporter activity. However, in the presence of Na+ and HCO3- during labeling both agents failed to inhibit cotransporter activity, indicating that these probes interact specifically with the cotransporter. In the presence of the substrates Na+ and HCO3-, PITC binds covalently to amino groups unprotected by substrates leaving the Na+/HCO3- cotransporter available for specific labeling with FITC. Addition of NaHCO3 to FITC-labeled proteoliposomes resulted in a concentration-dependent enhancement of the fluorescence signal which was inhibited by pretreatment with 4,4'-diisothiocyanostilbene 2',2-disulfonic acid (DIDS) prior to FITC labeling. SDS PAGE analysis of FITC-treated proteoliposomes showed the presence of two distinct fluorescent bands (approximate MW of 90 and 56 kD). In the presence of substrates, the fluorescence intensity of these bands was enhanced as confirmed by direct measurement of gel slice fluorescence. Thus, FITC detects conformational changes of the Na+/HCO3- cotransporter and labels proteins which may represent the cotransporter or components of this cotransporter. PMID:8051692

Stim, J; Bernardo, A A; Kear, F T; Qiu, Y Y; Arruda, J A

1994-05-01

274

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...

275

Technical procedures and staining methods for renal needle biopsies.  

PubMed

Procedures specifically developed for kidney needle biopsies are presented. These procedures may be applied to improve the slide quality of all small specimens. Fixation, processing, knife sharpening, cutting, and staining procedures used by the author are discussed. A modification of the Jones method for basement membranes is given. PMID:59547

Sugulas, M

1976-06-01

276

7 CFR 28.441 - Strict Middling Yellow Stained Color.  

Code of Federal Regulations, 2010 CFR

7 ? Agriculture ? 2 ? 2010-01-01 ? 2010-01-01 ? false ? Strict Middling Yellow Stained Color. ? 28.441 ? Section 28.441 ? Agriculture ? Regulations of the Department of Agriculture ? AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE ?...

2010-01-01

277

AUTOMATED STATISTICAL ANALYSIS OF COLOR-STAINED CELL IMAGES  

Microsoft Academic Search

A freely available collection of image analysis tools for color-stained biological cell images is presented. The anal- ysis platform includes a graphical user interface as well as a batch processing mode, and provides visualization pos- sibilities for statistical properties of cells. The tools fa- cilitate extraction of statistical information from, for ex- ample, cell-array images. The images to be analyzed

Jyrki Selinummi

278

Light microscope analysis of meiotic prophase chromosomes by silver staining  

Microsoft Academic Search

A method is described for the silver staining of the synaptonemal complex in surface-spread mammalian spermatocytes for light microscope examination. The method is quick, reliable, of broad applicability, and provides a means of making karyotype analysis at meiotic prophase. Many hundreds of suitable cells can be examined in an average preparation in a relatively short space of time. It has

Judith M. Fletcher

1979-01-01

279

Feulgen staining of intact plant tissues for confocal microscopy.  

PubMed

A method was developed to prepare plant structures for confocal laser scanning microscopy by combining Feulgen staining with pararosaniline and embedding in LR White(TM). This procedure preserves intact, delicate structures for three-dimensional imaging without loss from sectioning or squashing, and the slides can be viewed several times without serious photo-bleaching. PMID:9138536

Braselton, J P; Wilkinson, M J; Clulow, S A

1996-03-01

280

STAINING OF TISSUE SECTIONS FOR ELECTRON MICROSCOPY WITH HEAVY METALS  

Microsoft Academic Search

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

M. L. Watson

1958-01-01

281

A new thiacalix[4]arene-fluorescein based probe for detection of CN(-) and Cu(2+) ions and construction of a sequential logic circuit.  

PubMed

A new thiacalix[4]arene-fluorescein based fluorescent probe was synthesized, which shows a turn-on fluorescence response in the presence of CN(-) ions attributed to the nucleophilic addition of cyanide ions and the resulting cyanide adduct was used for the selective detection of copper ions. Furthermore, based on the fluorescence response a two input, one output, sequential logic circuit was constructed in the presence of CN(-) and Cu(2+) ions. PMID:25230713

Sharma, Neetu; Reja, Shahi Imam; Bhalla, Vandana; Kumar, Manoj

2014-10-01

282

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

E-print Network

standard UV illumination Compatible with SYPRO Ruby protein gel stain for detection of protein contaminants green fluorescence that is easy to visualize using a simple UV transilluminator. Contaminating proteins. They play a large role in protecting the bacterium from host defense mechanisms and antibiotics and trigger

Lebendiker, Mario

283

The compactness of populations of neutrophilic leucocytes as revealed by the new azure B-eosine stain, Pappenheim's stain and the peroxidase reaction  

Microsoft Academic Search

The new azure B-eosine stain shows a greater inhomogeneity of the granule population of human neutrophil leucocytes than Pappenheim's stain. This difference seems to be dependent on the number of azure granules within the single neutrophil.

H. Kurz; O. Leder

1984-01-01

284

Preparation of europium-quantum dots and europium-fluorescein composite nanoparticles available for ratiometric luminescent detection of metal ions.  

PubMed

The silica-encapsulated luminescent lanthanide nanoparticles have been developed for the selective tagging of a wide range of important targets in recent years, however, they are mainly limited to europium and terbium compounds. In this work, two types of europium-containing dual-luminophore silica nanoparticles, silica-encapsulated CdTe quantum dots (CdTe QDs)-BHHCT-Eu(3+) complex nanoparticles and BHHCT-Eu(3+) surface-bound silica-encapsulated fluorescein isothiocyanate (FITC) nanoparticles (BHHCT: 4, 4'-bis(1'', 1'', 1'', 2'', 2'', 3'', 3''-heptafluoro-4'', 6''-hexanedion-6''-yl)chlorosulfo-o-terphenyl), were successfully prepared using a water-in-oil (W/O) reverse microemulsion method. The results of transmission electron microscopy and luminescence spectroscopy characterizations indicate that the two types of nanoparticles are all monodisperse, spherical and uniform in size (approximately 50 nm in diameter), and have well-resolved and stable dual luminescence emission properties. The CdTe QDs-BHHCT-Eu(3+) nanoparticles can be excited at 365 nm to give dual-emission peaks at 535 and 610 nm, and the FITC-BHHCT-Eu(3+) nanoparticles can be excited at 335 nm to give dual-emission peaks at 515 and 610 nm. The luminescence response investigations of the nanoparticles to different metal ions indicate that the new nanoparticles can be used as ratiometric luminescent sensing probes for the selective detection of Cu(2+) and Fe(2+) ions, respectively. The performance of the nanoparticle probe for metal ion detection was investigated. PMID:20820091

Dong, Haitao; Liu, Yan; Wang, Dandan; Zhang, Wenzhu; Ye, Zhiqiang; Wang, Guilan; Yuan, Jingli

2010-10-01

285

High and Low Molecular Weight Fluorescein Isothiocyanate (FITC)-Dextrans to Assess Blood-Brain Barrier Disruption: Technical Considerations.  

PubMed

This note is to report how histological preparation techniques influence the extravasation pattern of the different molecular sizes of fluorescein isothiocyanate (FITC)-dextrans, typically used as markers for blood-brain barrier leakage. By using appropriate preparation methods, false negative results can be minimized. Wistar rats underwent a 2-h middle cerebral artery occlusion and magnetic resonance imaging. After the last imaging scan, Evans blue and FITC-dextrans of 4, 40, and 70 kDa molecular weight were injected. Different histological preparation methods were used. Sites of blood-brain barrier leakage were analyzed by fluorescence microscopy. Extravasation of Evans blue and high molecular FITC-dextrans (40 and 70 kDa) in the infarcted region could be detected with all preparation methods used. If exposed directly to saline, the signal intensity of these FITC-dextrans decreased. Extravasation of the 4-kDa low molecular weight FITC-dextran could only be detected using freshly frozen tissue sections. Preparations involving paraformaldehyde and sucrose resulted in the 4-kDa FITC-dextran dissolving in these reactants and being washed out, giving the false negative result of no extravasation. FITC-dextrans represent a valuable tool to characterize altered blood-brain barrier permeability in animal models. Diffusion and washout of low molecular weight FITC-dextran can be avoided by direct immobilization through immediate freezing of the tissue. This pitfall needs to be known to avoid the false impression that there was no extravasation of low molecular weight FITC-dextrans. PMID:21423333

Hoffmann, Angelika; Bredno, Jörg; Wendland, Michael; Derugin, Nikita; Ohara, Peter; Wintermark, Max

2011-03-01

286

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

E-print Network

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 with Coomassie Blue. However, the first silver staining protocols were not trouble-free. High

Paris-Sud XI, Université de

287

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

NASA Astrophysics Data System (ADS)

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

Shaw, Eric Max

288

Identifying neutrophils in H&E staining histology tissue images.  

PubMed

Identifying neutrophils lays a crucial foundation for diagnosing acute inflammation diseases. But, such computerized methods on the commonly used H&E staining histology tissue images are lacking, due to various inherent difficulties of identifying cells in such image modality and the challenge that a considerable portion of neutrophils do not have a "textbook" appearance. In this paper, we propose a new method for identifying neutrophils in H&E staining histology tissue images. We first segment the cells by applying iterative edge labeling, and then identify neutrophils based on the segmentation results by considering the "context" of each candidate cell constructed by a new Voronoi diagram of clusters of other neutrophils. We obtain good performance compared with two baseline algorithms we constructed, on clinical images collected from patients suspected of having inflammatory bowl diseases. PMID:25333103

Wang, Jiazhuo; MacKenzie, John D; Ramachandran, Rageshree; Chen, Danny Z

2014-01-01

289

Imaging port wine stains by fiber optical coherence tomography  

NASA Astrophysics Data System (ADS)

We develop a fiber optical coherence tomography (OCT) system in the clinical utility of imaging port wine stains (PWS). We use our OCT system on 41 patients with PWS to document the difference between PWS skin and contralateral normal skin. The system, which operates at 4 frames/s with axial and transverse resolutions of 10 and 9 ?m, respectively, in the skin tissue, can clearly distinguish the dilated dermal blood vessels from normal tissue. We present OCT images of patients with PWS and normal human skin. We obtain the structural parameters, including epidermal thickness and diameter and depth of dilated blood vessels. We demonstrate that OCT may be a useful tool for the noninvasive imaging of PWS. It may help determine the photosensitizer dose and laser parameters in photodynamic therapy for treating port wine stains.

Zhao, Shiyong; Gu, Ying; Xue, Ping; Guo, Jin; Shen, Tingmei; Wang, Tianshi; Huang, Naiyan; Zhang, Li; Qiu, Haixia; Yu, Xin; Wei, Xunbin

2010-05-01

290

Cement line staining in undecalcified thin sections of cortical bone  

NASA Technical Reports Server (NTRS)

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.

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

1990-01-01

291

Development of Cell Staining Technique for X-Ray Microscopy  

SciTech Connect

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.

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

292

Lectins stain cells differentially in the coral, Montipora capitata  

USGS Publications Warehouse

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.

Work, Thierry M.; Farah, Yael

2014-01-01

293

Selection of Ovine Oocytes by Brilliant Cresyl Blue Staining  

PubMed Central

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

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

2012-01-01

294

Nightguard vital bleaching: dark stains and long-term results.  

PubMed

Since its introduction to dentistry in 1989, nightguard vital bleaching has proven to be a simple and safe procedure for lightening discolored teeth. Efficacy of the technique is 98% for non-tetracycline-stained teeth, and with extended treatment time, tetracycline-stained teeth can be expected to lighten in 86% of cases. Satisfactory retention of the shade change without additional treatment can be expected in 63% of patients 3 years post-treatment and in at least 42% of patients at 7 years. Side effects are usually mild and transient, disappearing within days of treatment completion. Patients report that they are glad they went through the procedure and 98% recommend the procedure to a friend. PMID:11908344

Leonard, R H

2000-01-01

295

Identification of active fluorescence stained bacteria by Raman spectroscopy  

NASA Astrophysics Data System (ADS)

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.

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

2008-04-01

296

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.

297

'Catalysts' for polyacrylamide gel polymerization and detection of proteins by silver staining.  

PubMed

The crosslinker diacrylyl-piperazine produces polyacrylamide gels which display improved electrophoretic separation of proteins and better physical strength. It also produces gels with improved detection of proteins by ammoniacal silver staining by reducing the background. This reduced background provided us with an opportunity to investigate residual background staining caused by the catalytic reagents utilized in the polymerization of acrylamide gels. The commonly used catalyst system, tetramethyl-ethylenediamine and ammonium persulfate was shown to be responsible for the yellow staining background found after a prolonged development time with silver staining. An alternate catalyst system has been designed to decrease further the formation of this background staining. Dimethyl-piperazine or tetramethylethylenediamine, potassium or ammonium persulfate, and sodium thiosulfate are shown to provide for gels which have excellent mechanical and staining characteristics. These catalytic systems produce little background staining despite prolonged development time with the ammoniacal silver stain, and they reduce background staining with the dichromate silver stain. PMID:2484987

Hochstrasser, D F; Merril, C R

1988-01-01

298

The use of fluorescent-protein conjugates for staining brain capillaries in hypo- and hyperthermia (26-42 degrees).  

PubMed

The temperature dependency of cerebrocortical capillary diameter (CD), number perfused with fluorescent tracer (CN), and intercapillary distance (ICD) has been investigated for three body temperatures (26, 37, and 42 degrees), in order to analyze the influence of undesired cooling or warming which frequently occurs in animal experiments and humans (freezing, heat disposal, fever, etc.). The capillary bed (Wistar-Frömter rats, N = 92, ketaminxylazin anesthesia) was visualized with a double staining method using serum proteins coupled with either FITC (fluorescein isothiocyanate) or RB-200 (rhodamine-lissamin) injected ia immediately before decapitation. CD (6.1 +/- 0.3 micron, 37 degrees) increased during cooling by about 18%, during warming by about 13%. CN (313 +/- 83/mm2, 37 degrees) showed a 12% increase in response to temperature reduction and 18% after elevation. ICD was characterized by small insignificant changes of the mean values (44.2 +/- 5.5 micron, 37 degrees) at 26 degrees and 42 degrees. Mean cerebrocortical surface PO2 (sPO2) ranging between 15 and 22 mm Hg (37 degrees) increased slightly during warming or decreased during cooling. The sPO2 histogram showed a Gaussian-like shape in the range 0-40 mm Hg at low temperatures (26-34 degrees) and a left-shifted frequency distribution between 34-42 degrees. It was interesting to note that above 38 degrees sporadically high sPO2 values were registered in the range 40-80 mm Hg. Nevertheless, despite pronounced temperature variations, the net effect between O2 transport and consumption was balanced to such an extent that tissue anoxia was not detected within the rat cerebro-cortex. PMID:2448592

Metzger, H P; Brüggemann, H; Plewnia, A

1987-11-01

299

Ziehl-Neelsen Staining Technique Can Diagnose Paragonimiasis  

Microsoft Academic Search

BackgroundWe evaluated the Ziehl-Neelsen staining (ZNS) technique for the diagnosis of paragonimiasis in Laos and compared different modifications of the ZNS techniques.MethodologyWe applied the following approach: We (1) examined a paragonimiasis index case's sputum with wet film direct examination (WF) and ZNS; (2) re-examined stored ZNS slides from two provinces; (3) compared prospectively WF, ZNS, and formalin-ether concentration technique (FECT)

Günther Slesak; Saythong Inthalad; Phadsana Basy; Dalaphone Keomanivong; Ounheaun Phoutsavath; Somchaivang Khampoui; Aude Grosrenaud; Vincent Amstutz; Hubert Barennes; Yves Buisson; Peter Odermatt

2011-01-01

300

Nile red: a selective fluorescent stain for intracellular lipid droplets  

Microsoft Academic Search

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.

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

1985-01-01

301

Multicolor staining of root systems in pot culture  

Microsoft Academic Search

We have developed a method for staining the root systems of neighboring plants distinguishably in pot culture to facilitate studies of the interactions between plants. Pot soil was desiccated until the plant wilted, and then the shoot was cut and a dye solution (Fantasy) was pressure-injected into the roots at 0.05 MPa (gauge). All the roots, including fine roots of double-planted

Toshifumi Murakami; Satoshi Shimano; Satoshi Kaneda; Miyuki Nakajima; Yasufumi Urashima; Norikazu Miyoshi

2006-01-01

302

Alcian Blue Cartilage Staining Beatrice Jegalian & Eddy M. De Robertis  

E-print Network

Alcian Blue Cartilage Staining Beatrice Jegalian & Eddy M. De Robertis (Cell 71, 901-910, 1992) 1 will require more time) in 0.05% Alcian blue 8GX (Fisher) in 5% acetic acid. 6. Wash embryos in 5% acetic acid Alcian Blue 8 g NaCl 0.05% Alcian blue 8GX .2 g KCl 5% acetic acid 1.44 g Na2HPO4 water .24 g KH2PO4 800

De Robertis, Eddy M.

303

Response properties of stained monopolar cells in the honeybee lamina  

Microsoft Academic Search

1.Monopolar cells of the first visual ganglion, the lamina, of the bee were recorded from and stained intracellularly.2.Several different response types to pulses of spectral light were found. The most common response type hyperpolarized in a phasic-tonic fashion. The tonic hyperpolarizing response frequently decreased gradually, but in some cases increased with lasting illumination. Some cells also gave a transient response

John de Souza; Horst Hertel; Dora Fix Ventura; Randolf Menzel

1992-01-01

304

Revisit of imidazole-zinc reverse stain for protein polyacrylamide gel electrophoresis.  

PubMed

Imidazole-zinc reverse stain (ZN stain) is known for its high sensitivity, ease of use, and cost-effective feature. ZN stain is compatible to many experiments of which those are proteomics-related in particular. Here, we describe the ZN staining procedures and the subsequent procedures incorporated in detail, along with the improvements of setup in aspects of visualization and documentation for post-processing ZN-stained gel images. PMID:22585514

Chen, Han-Min

2012-01-01

305

Simultaneous staining with three fluorescent dyes of minute plankters on an agarose gel filter  

NASA Astrophysics Data System (ADS)

A new method, employing an agarose gel filter and triple staining with fluorescent dyes, was developed for observation and enumeration of planktonic microorganisms (0.2-20 ?m in size range) from a variety of niches in the marine ecosystem. Dansyl chloride was used to stain the cell-surface proteins, Calcofluor white was used to stain cellulose and chitin, and DAPI was used to stain DNA. The stained specimens also could be examined by transmission light microscopy.

Hara, Shigemitsu; Tanoue, Eiichiro

1989-11-01

306

The use of non-deparaffinized tissue sections for staining leprosy bacilli.  

PubMed

Reduced acid-fast staining of leprosy bacilli occurs during the dewaxing of paraffin sections by xylene and alcohols; the older and more decrepit bacilli being especially affected. By the use of non-deparaffinized sections, the leprosy bacilli which could not be stained with the usual carbol fuchsin are strongly stained. Moreover, non-deparafinized sections can be used for the periodic acid-carbol pararosanilin stain or methenamine silver stain for demonstrating mycobacteria. PMID:61950

Harada, K

1976-01-01

307

Post-processing application of chemical solutions for control of Listeria monocytogenes, cultured under different conditions, on commercial smoked sausage formulated with and without potassium lactate–sodium diacetate  

Microsoft Academic Search

This study evaluated post-processing chemical solutions for their antilisterial effects on commercial smoked sausage formulated with or without 1.5% potassium lactate plus 0.05% sodium diacetate, and contaminated (approximately 3–4 logcfu\\/cm2) with 10-strain composite Listeria monocytogenes inocula prepared under various conditions. Inoculated samples were left untreated, or were immersed (2min, 25±2°C) in solutions of acetic acid (2.5%), lactic acid (2.5%), potassium

Ifigenia Geornaras; Panagiotis N. Skandamis; Keith E. Belk; John A. Scanga; Patricia A. Kendall; Gary C. Smith; John N. Sofos

2006-01-01

308

En bloc staining with hydroquinone treatment for block face imaging.  

PubMed

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

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

2014-11-01

309

QZ1 and QZ2: Rapid, Reversible Quinoline-Derivatized Fluoresceins for Sensing Biological Zn(II)  

PubMed Central

QZ1, 2-[2-chloro-6-hydroxy-3-oxo-5-(quinolin-8-ylaminomethyl)-3H-xanthen-9-yl]benzoic acid, and QZ2, 2-[6-hydroxy-3-oxo-4,5-bis-(quinolin-8-ylaminomethyl)-3H-xanthen-9-yl]benzoic acid, two fluorescein-based dyes derivatized with 8-aminoquinoline, have been prepared and their photophysical, thermodynamic, and zinc-binding kinetic properties determined. Because of their low background fluorescence and highly emissive Zn(II) complexes, QZ1 and QZ2 have a large dynamic range, with 42- and 150-fold fluorescence enhancements upon Zn(II) coordination, respectively. These dyes have micromolar Kd values for Zn(II) and are selective for Zn(II) over biologically relevant concentrations of the alkali and alkaline earth metals. The Zn(II) complexes also fluoresce brightly in the presence of excess Mn(II), Fe(II), Co(II), Cd(II), and Hg(II), offering improved specificity for Zn(II) over di(2-picolyl)amine-based Zn(II) sensors. Stopped-flow kinetic investigations indicate that QZ1 and QZ2 bind Zn(II) with kon values of (3-4) × 106 M-1 s-1, compared to (6-8) × 105 M-1 s-1 for select ZP (Zinpyr) dyes, at 4.3 °C. Dissociation of Zn(II) from QZ1 and QZ2 occurs with koff values of 150 and 160 s-1, over 5 orders of magnitude larger than those for ZP probes, achieving reversibility on the biological (millisecond) time scale. Laser scanning confocal and two-photon microscopy studies reveal that QZ2 is cell-permeable and Zn(II)-responsive in vivo. Because of its weaker affinity for Zn(II), QZ2 responds to higher concentrations of intracellular Zn(II) than members of the ZP family, illustrating that binding affinity is an important parameter for Zn(II) detection in vivo. PMID:16316228

Nolan, Elizabeth M.; Jaworski, Jacek; Okamoto, Ken-Ichi; Hayashi, Yasunori; Sheng, Morgan

2005-01-01

310

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

PubMed

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

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

2012-04-30

311

Machine vision system for automated detection of stained pistachio nuts  

NASA Astrophysics Data System (ADS)

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

Pearson, Tom C.

1995-01-01

312

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

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

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

313

Photodynamic therapy of port wine stain: preliminary clinical studies  

NASA Astrophysics Data System (ADS)

The broad, long term objective of this work is the development of Photodynamic Therapy (PDT) for application in the clinical management of patients with port wine stain (PWS). PDT involves the use of an exogenous drug which is concentrated in a targeted tissue. When irradiated at wavelengths specifically absorbed by the drug, selective destruction of the targeted tissue, without the production of heat, occurs. The results of this preliminary study demonstrate in human PWS patients that a photosensitizer, such as PHOTOFRINR, activated by red light at the appropriate therapeutic wavelength, can cause destruction of subsurface blood vessels in the skin with a high degree of specificity, and further study appears warranted.

Nelson, J. Stuart

1993-07-01

314

Ring stains in the presence of electromagnetohydrodynamic interactions  

NASA Astrophysics Data System (ADS)

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.

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

2012-11-01

315

Interphase ribosomal RNA cistron staining in chronic myeloid leukaemia  

PubMed Central

Aim—To evaluate the haemopoietic function of bone marrow blood forming cells in human chronic myeloid leukaemia (CML) by means of silver staining of nucleolar organiser region (AgNOR). Methods—Nucleoli were investigated in bone marrow blast cells and in erythroid, granulocytic, and megakaryocytic cells from 10 haematologically healthy subjects and from 26 patients with chronic myeloid leukemia (17 in benign phase, nine with blast crisis). The investigation was done before treatment, by means of a one step silver staining method. In every case 50 to 100 blasts, promyelocytes, myelocytes, immature (pronormoblastic and basophilic normoblastic) and mature (polychromatic normoblastic) erythroid elements, and megakaryocytes were evaluated for the mean numbers of nucleoli and for the average number of AgNORs per nucleus. Student's t test was used to compare the patient and control groups. Other statistical analyses were carried out by means of the computer assisted “HEMA” system. Results—Compared with controls, activation of NORs was noticed only in CML blasts, while there was a decrease in NORs in the erythroid elements, promyelocytes, and megakaryocytes. The AgNOR score of polychromatic normoblasts and megakaryocytes started to decrease in the benign stage of CML, whereas a similar decrease in pronormoblasts, basophilic normoblasts, and promyelocytes was detected only in patients with CML blast crisis. Conclusions—The loss of AgNOR sites in cell series in CML may be related to intrinsic defects in their proliferation. PMID:16696018

Mamaev, N N; Salogub, G N; Koloskov, A V

1995-01-01

316

Interphase ribosomal RNA cistron staining in chronic myeloid leukaemia.  

PubMed

Aim-To evaluate the haemopoietic function of bone marrow blood forming cells in human chronic myeloid leukaemia (CML) by means of silver staining of nucleolar organiser region (AgNOR).Methods-Nucleoli were investigated in bone marrow blast cells and in erythroid, granulocytic, and megakaryocytic cells from 10 haematologically healthy subjects and from 26 patients with chronic myeloid leukemia (17 in benign phase, nine with blast crisis). The investigation was done before treatment, by means of a one step silver staining method. In every case 50 to 100 blasts, promyelocytes, myelocytes, immature (pronormoblastic and basophilic normoblastic) and mature (polychromatic normoblastic) erythroid elements, and megakaryocytes were evaluated for the mean numbers of nucleoli and for the average number of AgNORs per nucleus. Student's t test was used to compare the patient and control groups. Other statistical analyses were carried out by means of the computer assisted "HEMA" system.Results-Compared with controls, activation of NORs was noticed only in CML blasts, while there was a decrease in NORs in the erythroid elements, promyelocytes, and megakaryocytes. The AgNOR score of polychromatic normoblasts and megakaryocytes started to decrease in the benign stage of CML, whereas a similar decrease in pronormoblasts, basophilic normoblasts, and promyelocytes was detected only in patients with CML blast crisis.Conclusions-The loss of AgNOR sites in cell series in CML may be related to intrinsic defects in their proliferation. PMID:16696018

Mamaev, N N; Salogub, G N; Koloskov, A V

1995-10-01

317

The Golgi Stain: invention, diffusion and impact on neurosciences.  

PubMed

The black reaction, invented in 1873 by Camillo Golgi (1843-1926, was the first technique to reveal neurons in their entirety, i.e. with all their processes. This important development passed unnoticed at first and only received wide international attention after a long delay. The Golgi stain was widely employed for almost thirty years and was directly responsible for major advances in our knowledge of the microscopic anatomy of the nervous system, as well as in other fields of study. In the hands of other researchers, the black reaction provided vital evidence that helped to establish the neuron theory. The Golgi stain was almost forgotten in the period between the two World Wars, but the introduction of the electron microscope to neurocytological resarch revived its use around the middle of the twentieth century. Today, the black reaction is still used extensively not only in combination with electron microscopy, but also as an autonomous technique in studies on the evolution, ontogeny, and organization of the nervous system. PMID:11624294

Pannese, E

1999-08-01

318

Responses of acid-stressed Salmonella Typhimurium in broth and chicken patties to subsequent antimicrobial stress with epsilon-polylysine and combined potassium lactate and sodium diacetate.  

PubMed

We investigated the growth kinetics and morphological changes in acid-stressed Salmonella Typhimurium as well as the antimicrobial effects of epsilon-polylysine (SAVE-ORY GL610) and combined potassium lactate (PL) and sodium diacetate (SDA) (PURASAL Opti.Form PD Plus) on acid-stressed S. Typhimurium. Exposure to 0.5% acetic or lactic acid injured over 90% of the S. Typhimurium population. Although the lag time of the injured S. Typhimurium was extended, the injured cells were recovered at 10 degrees C and 24 degrees C, indicating a risk of using 10 degrees C as a storage temperature. Additionally, 4.5% PL/SDA mixture or 2% epsilon-polylysine completely inhibited the growth of acid-stressed S. Typhimurium in broth at 10, 24, or 35 degrees C. Although 3% PL/SDA mixture inhibited the growth of lactic acid-stressed S. Typhimurium at 10 degrees C, it did not inhibit the growth of unstressed S. Typhimurium at the same temperature. This finding indicates a different antimicrobial effect due to the physiological status of the pathogen. Furthermore, acid-stressed S. Typhimurium was not resistant to epsilon-polylysine or the PL/SDA mixture, although the antimicrobial effect of these compounds was enhanced at a lower storage temperature. TEM analysis revealed that most of the stressed cells lost their cellular integrity and membranes partially. Both dead and doubling cells were observed after recovery at 30 degrees C for 12 h. The addition of 2% epsilon-polylysine or 4.5% PL/SDA mixture resulted in the collapse of the structure of S. Typhimurium cells and cytoplasmic materials being released. These results provide valuable information regarding the morphological and physiological responses of acid-stressed S. Typhimurium cells in broth and chicken patties followed by antimicrobial stress with epsilon-polylysine or PL/SDA mixture. PMID:19465242

Jung, Y J; Min, K J; Yoon, K S

2009-08-01

319

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

SciTech Connect

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

Rastogi, Rajesh P. [Department of Biology, Friedrich-Alexander University Erlangen-Nuremberg, Staudtstrasse 5, D-91058 Erlangen (Germany) [Department of Biology, Friedrich-Alexander University Erlangen-Nuremberg, Staudtstrasse 5, D-91058 Erlangen (Germany); Laboratory of Photobiology and Molecular Microbiology, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005 (India); Singh, Shailendra P.; Haeder, Donat-P. [Department of Biology, Friedrich-Alexander University Erlangen-Nuremberg, Staudtstrasse 5, D-91058 Erlangen (Germany)] [Department of Biology, Friedrich-Alexander University Erlangen-Nuremberg, Staudtstrasse 5, D-91058 Erlangen (Germany); Sinha, Rajeshwar P., E-mail: r.p.sinha@gmx.net [Laboratory of Photobiology and Molecular Microbiology, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi 221005 (India)

2010-07-02

320

A Procedure for Laundering Oil-Stained Durable Press Utility Uniforms.  

National Technical Information Service (NTIS)

During the summer of 1973, durable press, cotton/polyester utility uniforms were field tested (TECOM) at Fort Benning, Georgia. Tests showed that the prescribed laundry procedures did not remove oil and grease stains from these garments. Stain removal was...

H. T. Skerritt

1975-01-01

321

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

E-print Network

stains can detect 48 ng of protein per minigel band, higher sensitivity than Coomassie® brilliant blue) Coomassie brilliant blue (CBB) stain according to standard proto- cols. The SYPRO dyestained gels were

Lebendiker, Mario

322

Development and Testing of a Rapid Multiplex Assay for the Identification of Biological Stains.  

National Technical Information Service (NTIS)

While DNA profiling makes it possible to individualize biological stains, the identification of the stain itself can present forensic serologists with a significant challenge. Current antibody- and enzyme activity-based assays used by forensic practitione...

K. M. Legg

2013-01-01

323

Visualization of nucleolar organizer regions in mammalian chromosomes using silver staining  

Microsoft Academic Search

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

Carll Goodpasture; Stephen E. Bloom

1975-01-01

324

Nucleic acid stains as indicators of Giardia muris viability following cyst inactivation  

Microsoft Academic Search

A reliable viability assay for Giardia is required for the development of disinfection process design criteria and pathogen monitoring by water treatment utilities. Surveys of single-staining nucleic acid dyes (stain dead parasites only), and double-staining vital dye kits from Molecular Probes (stain live and dead parasites) were conducted to assess the viability of untreated, heat-killed, and chemically inactivated Giardia muris

R. Taghi-Kilani; L. L. Gyürék; P. J. Millard; G. R. Finch; M. Belosevic

1996-01-01

325

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

Microsoft Academic Search

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

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

2000-01-01

326

Are routine iron stains on bone marrow trephine biopsy specimens necessary?  

Microsoft Academic Search

Aims: To determine the role of Perls’ staining in bone marrow trephine biopsy sections.Methods: The haemosiderin content of 155 Perls’ stained, formic acid decalcified trephine biopsy sections was assessed and compared with Perls’ stained aspirate samples in 105 cases and haematoxylin and eosin (H&E) stained biopsy sections in all cases.Results: An evaluable aspirate film with positive iron or at least

S E Stuart-Smith; D A Hughes; B J Bain

2005-01-01

327

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

E-print Network

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

Brooks, Stephen

328

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

Microsoft Academic Search

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

S. E. Bloom; C. Goodpasture

1976-01-01

329

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

E-print Network

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

Løw, Erik

330

AN INVESTIGATION INTO THE STAINING PROBLEM IN WESTERN HEMLOCK (A TEST CASE)  

Microsoft Academic Search

The mill grader reported that a visual inspection revealed that from 2% to 25% of the pieces had some stains, and the stains ranged in color from brown and gray to even a light orange. With many differing opinions expressed as to the cause of the stains, the mill manager decided to assemble a team to investigate the cause of

Sita Millar

331

Numerical modeling of spray cooling-assisted dermatologic laser surgery for treatment of port wine stains  

E-print Network

to the epidermis during dermatologic laser surgery (DLS) for removal of port wine stain (PWS) birthmarks stains Walfre Francoa,b, Rong Zhangb, J. Stuart Nelsonb, and Guillermo Aguilara,b aDept. of Mechanical) to treat vascu- lar superficial lesions and abnormalities, such as port wine stains (PWS) birthmarks

Aguilar, Guillermo

332

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

Microsoft Academic Search

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

Gang Wang; Dongmei Yan; Yang Yang

2010-01-01

333

Silver Staining SDS Gels Remove the stacking gel before the first fixative step.  

E-print Network

37 Silver Staining SDS Gels · Remove the stacking gel before the first fixative step. · Do, for frequent silver staining: make up solutions as 10X stocks ahead of time. 1. Fix in 50% methanol, 10% acetic seconds. Repeat the change of developer solution once again. Agitate gently until the desired staining

Aris, John P.

334

bleachingThe chemical stain removal properties of 'whitening' toothpaste products: studies in vitro  

Microsoft Academic Search

Background A considerable number of toothpastes are available as tooth whitening products. Most appear to contain ingredients that might remove extrinsic stains rather than change natural tooth colour. Extrinsic stain removal could be achieved by physical or chemical means.Aim The purpose of this study was to measure the chemical stain removal properties of a range of whitening toothpaste products and

N Sharif; E MacDonald; J Hughes; R G Newcombe; M Addy

2000-01-01

335

Effectiveness and Mechanisms of Action of Whitening Dentifrices on Enamel Extrinsic Stains. Salem Alshara1  

E-print Network

of Dentistry. Whitening dentifrices utilize different approaches for stain removal and/or prevention, includingEffectiveness and Mechanisms of Action of Whitening Dentifrices on Enamel Extrinsic Stains. Salem six bovine enamel specimens (10x10mm) were prepared and partially stained. They were assigned to 8

Zhou, Yaoqi

336

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

NASA Astrophysics Data System (ADS)

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

Bautista, Pinky A.; Yagi, Yukako

2012-05-01

337

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

E-print Network

photographic filter and Polaroid 667 black-and-white print film. The CBB- and silver-stained gels were CoomassieTM Brilliant Blue and as sensitive as silver staining " Rapid. Staining complete in (Figure 2) and, with the correct filters, work well with CCD camera archiving systems. SYPRO Orange

Lebendiker, Mario

338

The Pattern of Ocular Dominance Columns in Macaque Visual Cortex Revealed by a Reduced Silver Stain  

E-print Network

The Pattern of Ocular Dominance Columns in Macaque Visual Cortex Revealed by a Reduced Silver Stain- face, were seen in tangential sections stained with a reduced silver method for normal fibers and were fixed, sectioned tangentially and stained with the silver method. All the lesions- a total of 12 -fell

Hubel, David

339

Phos-toolsTM Phos-tagTM 540 Phosphoprotein Blot Stain  

E-print Network

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

Lebendiker, Mario

340

Human intestinal mucosal mast cells: evaluation of fixation and staining techniques  

Microsoft Academic Search

The staining properties of tissue mast cells are influenced by the method of fixation. Differences in fixation and staining techniques may explain the contradictory results in the published reports on the number of human mucosal mast cells (MMC) in the gastrointestinal mucosa in health and disease. We have examined the influence of fixatives on the staining properties of human MMC

S Strobel; H R Miller; A Ferguson

1981-01-01

341

A comparative study of quantitative stains for DNA in image cytometry.  

PubMed

In this study we examined the reproducibility of several stains used to measure nuclear DNA by image cytometry. The specimens were touch preparations of liver and testis from mouse and liver, intestine and brain from rat, fixed in either neutral formalin or Carnoy's solution. The tested stains included four Feulgen methods (pararosaniline, azure-A, thionin and acriflavine), the gallocyanine-chromalum stain and two fluorescent stains (acridine orange and propidium iodide). Absorbance measurements employed a video image analysis system; fluorescence measurements were from a scanning microspectrophotometer. The acriflavine-Feulgen stain was analyzed for both absorbance and fluorescence. All seven stains were quantitative for DNA and gave reproducible results. The absorbance measurements had a lower coefficient of variation (CV) than the fluorescence values. In a nested analysis of variance of the pararosaniline Feulgen stains, cell-to-cell variability accounted for 67% of the total variance; slide-to-slide, 9%; and batch-to-batch, 24%. These values did not change significantly when the staining was performed in an automatic staining machine. For DNA analysis using image cytometry, we conclude that the Feulgen staining technique is the most useful. In particular, acriflavine-Feulgen-stained cells fixed in Carnoy's fluid give the least variation between measurement values and the most accurate ratios between the separate ploidy groups. For fluorescence cytometry we recommend Carnoy's fixation and the acriflavine-Feulgen stain because of its narrow CV as compared to acridine orange and propidium iodide. PMID:1718295

Mikel, U V; Becker, R L

1991-08-01

342

Fat tissue staining and photodynamic/photothermal effects  

NASA Astrophysics Data System (ADS)

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.

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

2010-02-01

343

Sizing of single fluorescently stained DNA fragments by scanning microscopy  

PubMed Central

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

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

2003-01-01

344

Conjugates of a Photoactivated Rhodamine with Biopolymers for Cell Staining  

PubMed Central

Conjugates of the photoactivated rhodamine dyes with biopolymers (proteins, polysaccharides, and nucleic acids) are important tools for microscopic investigation of biological tissue. In this study, a precursor of the photoactivated fluorescent dye (PFD) has been successfully used for staining of numerous mammalian cells lines and for conjugate formation with chitosan (“Chitosan-PFD”) and histone H1 (“Histone H1.3-PFD”). The intensive fluorescence has been observed after photoactivation of these conjugates inside cells (A431, HaCaT, HEK239, HBL-100, and MDCK). Developed procedures and obtained data are important for further application of novel precursors of fluorescent dyes (“caged” dyes) for microscopic probing of biological objects. Thus, the synthesized “Chitosan-PFD” and “Histone H1-PFD” have been successfully applied in this study for intracellular transport visualization by fluorescent microscopy. PMID:25383365

Zaitsev, Sergei Yu.; Shaposhnikov, Mikhail N.; Solovyeva, Daria O.; Solovyeva, Valeria V.; Rizvanov, Albert A.

2014-01-01

345

Early morphea mimicking acquired port-wine stain.  

PubMed

We report the case of a 2.5-year-old girl with linear morphea initially diagnosed as an acquired port-wine stain (PWS). She underwent three treatments to the right face using the pulsed dye laser (PDL) before sclerotic changes were observed and the correct diagnosis was confirmed with histopathology. Treatment using the PDL reduced the skin erythema but did not prevent subsequent sclerosis. The sclerosis became most prominent superior to the patient's right ear in an area not treated using the laser. A review of the English-language medical literature identified no cases of morphea triggered using a PDL, but there were several reports of early morphea misdiagnosed as an acquired PWS. Briefly, we review those cases, as well as morphea subtypes, and comment on how the pathophysiology of morphea may lend itself to an early underrecognized inflammatory presentation, delaying diagnosis. PMID:23627630

Pickert, Amanda J; Carpentieri, David; Price, Harper; Hansen, Ronald C

2014-01-01

346

A study to evaluate the efficacy of xylene-free hematoxylin and eosin staining procedure as compared to the conventional hematoxylin and eosin staining: An experimental study  

PubMed Central

Context: Use of diluted dish washing solution (DWS) has been experimented successfully as a substitute for xylene to deparaffinize tissue sections during hematoxylin and eosin (H and E) staining. Aims: (1) Test the hypothesis that xylene- and methanol-free sections (XMF) deparaffinized with diluted DWS are better than or at par with conventional H and E sections. (2) To compare the efficacy of xylene-free sections with the conventional H and E sections. Settings and Design: Single blinded experimental study. Materials and Methods: Sixty paraffin blocks were considered. One section was stained with conventional H and E method (Group A) and the other with XMF H and E (Group B). Slides were scored for parameters; nuclear staining, cytoplasmic staining (adequate = score1, inadequate = score0), uniformity, clarity, crispness (present = score1, absent = score0). Score >/= 2 was inadequate for diagnosis and 3-5 was adequate for diagnosis. Statistical analysis used: Z test. Results: Adequate nuclear staining, 96.66% sections in group A and 98.33% in Group B (Z = 0.59, P>0.05); adequate cytoplasmic staining, 93.33% in group A and 83.33% in Group B (Z = 1.97, P<0.05); uniform staining, 70% in group A, 50% in group B (Z = 1.94, P<0.05), clarity present in 85% of group A, 88.33% of group B sections (Z = 0.27, P>0.05), crisp staining in 76.66% in group A and 83.33% in Group B (Z = 1.98, P<0.05), 88.33% Group A sections stained adequately for diagnosis as compared with 90% in Group B (Z = 0.17, P>0.05). Conclusion: Xylene- and methanol-free H and E staining is a better alternative to the conventional H and E staining procedure. PMID:22529574

Ankle, Madhuri R; Joshi, Priya S

2011-01-01

347

Optimized Negative-Staining Electron Microscopy for Lipoprotein Studies  

PubMed Central

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

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

2012-01-01

348

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

Microsoft Academic Search

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

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

1995-01-01

349

PREFERENTIAL STAINING OF NUCLEIC ACID-CONTAINING STRUCTURES FOR ELECTRON MICROSCOPY  

PubMed Central

Oriented fibres of extracted nucleohistone were employed as test material in a study of satisfactory fixation, embedding, and staining methods for structures containing a high proportion of nucleic acid. Fixation in buffered osmium tetroxide solution at pH 6, containing 10-2 M Ca++, and embedding in Araldite enabled sections of the fibres to be cut in which the orientation was well preserved. These could be strongly stained in 2 per cent aqueous uranyl acetate, and showed considerable fine structure. Certain regions in the nuclei of whole thymus tissue could also be strongly stained by the same procedure, and were identical with the regions stained by the Feulgen procedure in adjacent sections. Moreover, purified DNA was found to take up almost its own dry weight of uranyl acetate from 2 per cent aqueous solution. Strongest staining of whole tissue was obtained with very short fixation times-5 minutes or so at 0°C. Particularly intense staining was obtained when such tissue stained in uranyl acetate was further stained with lead hydroxide. Although the patterns of staining by lead hydroxide alone and by uranyl acetate were similar in tissues fixed for longer times (½ hour to 2 hours, at 0°C or 20°C), in briefly fixed material the DNA-containing regions appeared relatively unstained by lead hydroxide alone, whilst often there was appreciable staining of RNA-containing structures. Observations on the staining of some viruses by similar techniques are also described. PMID:14450292

Huxley, H. E.; Zubay, G.

1961-01-01

350

Digital staining for multispectral images of pathological tissue specimens based on combined classification of spectral transmittance.  

PubMed

In this study, the digital transformation (digital staining) of the 16-band multispectral image of a hematoxylin and eosin (HE) stained pathological specimen to its Masson's trichrome (MT) stained counterpart is addressed. The digital staining procedure involves the classification of the various H&E-stained tissue components and then the transformation of their transmittance spectra to their equivalent MT-stained transmittance configurations. Combination of transmittance classifiers were designed to classify the various tissue components found in the multispectral images of an HE-stained specimen, e.g. nucleus, cytoplasm, red blood cell (RBC), fibrosis, etc.; while pseudo-inverse method was used to obtain the transformation matrices that would translate the transmittance spectra of the classified HE-stained multispectral pixels to their MT-stained configurations. To generate the digitally stained image, weighting factors, which were based on the classifiers beliefs, were introduced to the generated transformation matrices. Initial results of our experiments on liver specimens show the viability of multispectral imaging (MSI) to implement a digital staining framework in the pathological context. PMID:16269238

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

2005-12-01

351

Immunopathological stain of lipoarabinomannan-B (LAM-B) for diagnosis of leprosy.  

PubMed

We developed an immunopathological staining of LAM-B antigen in formalin-fixed paraffin-embedded tissues, and compared it with, PGL-I immunostaining, Fite Faraco's stain and periodic acid carbol pararosaniline (PACPR) stain. Out of the total 28 leprosy cases, 27 were positive to LAM-B immunostaining while 23 were positive to PGL-I stain. Fite's stain was positive in 21 cases while PACPR stain was positive in 24 cases. In scrofuloderma, LAM-B antigen was observed only in the granuloma while no other positive findings were noted with other stains. Normal skin did not give any positive findings with any of the stains. Other dermatoses showed no positive findings to any of the stains tested. LAM-B staining was observed in the nerve even in the absence of bacilli in leprosy tissues. Presence of LAM-B in the cutaneous nerves is helpful in discriminating leprosy from other mycobacterioses. Considering the high sensitivity of LAM-B and the predilection of M. leprae for the nerves, we concluded that LAM-B staining can be a useful new tool in the prompt diagnosis of leprosy, especially in suspected or early cases. PMID:7693642

Butt, K I; Kawatsu, K; Wang, T; Maeda, Y; Izumi, S

1993-03-01

352

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

PubMed

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

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

2014-04-01

353

Evaluating the growth of Listeria monocytogenes in refrigerated ready-to-eat frankfurters: influence of strain, temperature, packaging, lactate and diacetate, and background microflora.  

PubMed

This research was conducted to study the growth of Listeria monocytogenes inoculated on frankfurters stored at different conditions as a basis for a safety-based consume by shelf life date label. Three L. monocytogenes strains were separately inoculated at 10 to 20 CFU/cm2 onto frankfurters that were previously formulated with or without high pressure and with or without added 2% potassium lactate (PL) and 0.2% sodium diacetate (SD). Inoculated frankfurters were air or vacuum packaged; stored at 4, 8, or 12 degrees C; and L. monocytogenes and psychrotrophic plate counts were determined for 90, 60, and 45 days, respectively, or until the stationary phase was reached. The data (log CFU per square centimeter versus time) were fitted using the Baranyi-Roberts model to determine maximum growth rates and lag-phase time. The maximum growth rates and the lag time under each growth condition were used to calculate the time to reach 100-fold the initial Listeria population. In frankfurters lacking PL and SD, the count of all strains increased by 2 log after 18 to 50 days at 4 degrees C and 4 to 13 days at 8 degrees C. The growth was inhibited at 4 and 8 degrees C in frankfurters containing PL and SD, but one ribotype was capable of growing, with the time to reach 100-fold the initial Listeria population ranging from 19 to 35 days at 12 degrees C. In most cases, the time to reach 100-fold the initial Listeria population of L. monocytogenes was significantly longer in vacuum-packaged frankfurters as compared with air-packaged samples. Inclusion of PL and SD also inhibited the growth of psychrotrophs, but at all temperatures the psychrotrophic plate counts were greater than 4 log CFU/cm2 at the end of the experiments. These results indicated that despite the use of antimicrobials, certain L. monocytogenes strains could be capable of growing under storage-abuse conditions. Growth kinetics data could be useful for establishing a shelf life date label protocol under different handling scenarios. PMID:18810864

Pal, Amit; Labuza, Theodore P; Diez-Gonzalez, Francisco

2008-09-01

354

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

NASA Astrophysics Data System (ADS)

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

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

2005-02-01

355

A nest of structures in dynamics of cellulose diacetate in N,N-dimethylacetamide in quiescent solution state studied by dynamic light scattering  

NASA Astrophysics Data System (ADS)

In the quiescent state, dynamic light scattering measurements were performed for a polysaccharide, cellulose diacetate (CDA, the degree of substitution 2.40), in a liquid-crystalline promoting solvent N, N-dimethylacetamide (DMAc) in the range from dilute to crossover concentrations. Three modes of motions were detected simultaneously over all concentrations measured. They are in a nest of structures, which are caused by long-range interactions acting between OH groups of glucose units in a highly dielectric-constant solvent DMAc. Denoting these modes as Mode I, II, and III from fast to slow motions, the decay rates ? of each mode showed the squared scattering-vector dependence at lower angles, i.e., the diffusion nature. In dilute solution, fast Mode I is the translational diffusion of a single CDA chain and others represent the dissipation of locally associated CDA clouds which grow by the interactions specified above. Mode II is related to a coarse cloud formed in a limited time but spread over a wide space A, while Mode III is related to a dense cloud created with a very small space B inside >A. The tentative increase in CDA concentrations in the spaces A and B relaxes to the level of bulk concentration with a large decay rate ?II for Mode II and a small ?III for Mode III. With the increase of c, the number of A and the bulk concentration increase, and the decay rate ?III for dissipating B becomes smaller than that in dilute solution. Above the crossover concentration c*, however, the space A becomes close each other and almost all the CDA are packed in A. Then fast Mode I represents the relaxation of concentration fluctuations, or cooperative diffusion, with the length scale as small as a single CDA chain, ?I. Mode II represents another cooperative diffusion, which is originated by the relaxation of concentration fluctuations effective only in A of the correlation length ?II. Mode III is related to the time-space fluctuation in the distribution of A, which emerges as another cooperative diffusion of much larger correlation length ?III. Mode II was contaminated by a self-diffusionlike (SF) motion, though both were separated into two branches in more concentrated regions. The SF mode is assigned to a reptation motion of CDA, which can be described by the constrained translational motion of a stiff chain in the tube with interacting wall. The dynamical feature of these modes of motions are influenced strongly by strength and nature of long-range interactions between unsubstituted OH groups in glucose residues, or the hydrogen bonds in solvents of high electronegativity.

Kawanishi, Hiroyuki; Tsunashima, Yoshisuke; Horii, Fumitaka

1998-12-01

356

Quantitative risk assessment for Listeria monocytogenes in selected categories of deli meats: impact of lactate and diacetate on listeriosis cases and deaths.  

PubMed

Foodborne disease associated with consumption of ready-to-eat foods contaminated with Listeria monocytogenes represents a considerable pubic health concern. In a risk assessment published in 2003, the U.S. Food and Drug Administration and the U.S. Food Safety and Inspection Service estimated that about 90% of human listeriosis cases in the United States are caused by consumption of contaminated deli meats. In this risk assessment, all deli meats were grouped into one of 23 categories of ready-to-eat foods, and only the postretail growth of L. monocytogenes was considered. To provide an improved risk assessment for L. monocytogenes in deli meats, we developed a revised risk assessment that (i) models risk for three subcategories of deli meats (i.e., ham, turkey, and roast beef) and (ii) models L. monocytogenes contamination and growth from production to consumption while considering subcategory-specific growth kinetics parameters (i.e., lag phase and exponential growth rate). This model also was used to assess how reformulation of the chosen deli meat subcategories with L. monocytogenes growth inhibitors (i.e., lactate and diacetate) would impact the number of human listeriosis cases. Use of product-specific growth parameters demonstrated how certain deli meat categories differ in the relative risk of causing listeriosis; products that support more rapid growth and have reduced lag phases (e.g., turkey) represent a higher risk. Although reformulation of deli meats with growth inhibitors was estimated to reduce by about 2.5- to 7.8-fold the number of human listeriosis cases linked to a given deli meat subcategory and thus would reduce the overall risk of human listeriosis, even with reformulation deli meats would still cause a considerable number of human listeriosis cases. A combination of strategies is thus needed to provide continued reduction of these cases. Risk assessment models such as that described here will be critical for evaluation of different control approaches and to help define the combinations of control strategies that will have the greatest impact on public health. PMID:19517724

Pradhan, Abani K; Ivanek, Renata; Gröhn, Yrjö T; Geornaras, Ifigenia; Sofos, John N; Wiedmann, Martin

2009-05-01

357

Development of a stained cell nuclei counting system  

NASA Astrophysics Data System (ADS)

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.

Timilsina, Niranjan; Moffatt, Christopher; Okada, Kazunori

2011-03-01

358

p63 immunohistochemical staining is limited in soft tissue tumors.  

PubMed

p63 is a p53 homolog that is expressed in various normal epithelial tissues and epithelial malignancies. Its expression in mesenchymal lesions has not been examined in depth; therefore, we studied p63 expression by immunohistochemical analysis in 650 soft tissue tumors. We found that p63 expression is limited in soft tissue tumors. The majority of tumors studied were p63-, including all cases of angiosarcoma, lipomatous neoplasms, dermatofibrosarcoma protuberans, solitary fibrous tumor, schwannoma, neurofibroma, gastrointestinal stromal tumor, and leiomyosarcoma. Nuclear p63 reactivity was found in a subset of soft tissue myoepithelioma and myoepithelial carcinoma of soft tissue, cellular neurothekeoma, soft tissue perineurioma, Ewing sarcoma/peripheral neuroectodermal tumor, diffuse-type giant cell tumor, and giant cell tumor of soft parts. Infrequent, weak, or focal p63-staining patterns were observed in low-grade fibromyxoid sarcoma, malignant peripheral nerve sheath tumor, extraskeletal myxoid chondrosarcoma, myxofibrosarcoma, proximal-type epithelioid sarcoma, synovial sarcoma, embryonal rhabdomyosarcoma, desmoplastic small round cell tumor, atypical fibroxanthoma, and spindle cell melanoma. Absent p63 expression is typical for most soft tissue tumors, including most (but not all) that would be in the differential diagnosis of spindle cell squamous carcinoma. PMID:22031315

Jo, Vickie Y; Fletcher, Christopher D M

2011-11-01

359

The challenges of analysing blood stains with hyperspectral imaging  

NASA Astrophysics Data System (ADS)

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.

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

2014-06-01

360

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

NASA Astrophysics Data System (ADS)

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.

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

2013-05-01

361

Is the gram stain useful in the microbiologic diagnosis of VAP? A meta-analysis.  

PubMed

In a meta-analysis examining respiratory specimen Gram stain for diagnosis of ventilator-associated pneumonia, absence of bacteria on Gram stain had a high negative predictive value, but a positive Gram stain correlated poorly with organisms recovered in culture. Rapid and accurate diagnosis of ventilator-associated pneumonia (VAP) is a major challenge and no generally accepted gold standard exists for VAP diagnosis. We conducted a meta-analysis to examine the role of respiratory specimen Gram stain to diagnose VAP, and the correlation with final culture results. In 21 studies, pooled sensitivity of Gram stain for VAP was 0.79 (95% confidence interval [CI], .77-0.81; P < .0001) and specificity was 0.75 (95% CI, .73-.78; P < .0001). Negative predictive value of Gram stain for a VAP prevalence of 20%-30% was 91%, suggesting that VAP is unlikely with a negative Gram stain but the positive predictive value of Gram stain was only 40%. Pooled kappa was 0.42 for gram-positive organisms and 0.34 for gram-negative organisms, suggesting fair concordance between organisms on Gram stain and recovery by culture. Therefore, a positive Gram stain should not be used to narrow anti-infective therapy until culture results become available. PMID:22677711

O'Horo, John C; Thompson, Deb; Safdar, Nasia

2012-08-01

362

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

PubMed

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

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

2012-11-30

363

pH-dependent Si-fluorescein hypochlorous acid fluorescent probe: spirocycle ring-opening and excess hypochlorous acid-induced chlorination.  

PubMed

We report the synthesis and characterization of a fluorescent probe (Hypo-SiF) designed for the detection of hypochlorous acid (HOCl) using a silicon analogue of fluorescein (SiF). The probe is regulated in an "off-on" fashion by a highly selective thioether spirocyclic nonfluorescent structure that opens to form a mixture of fluorescent products in the presence of HOCl. Over a range of pH values, the probe reacts with a stoichiometric amount of HOCl, resulting in a mixture of two pH-dependent fluorescent species, a SiF disulfide product and a SiF sulfonate product. The unique colorimetric properties of the individual SiF fluorophores were utilized to perform simultaneous detection of HOCl and pH. When an excess of HOCl is present, the SiF fluorophores become chlorinated, via an intermediate halohydrin, resulting in a more pH independent and red-shifted fluorophore. PMID:23889259

Best, Quinn A; Sattenapally, Narsimha; Dyer, Daniel J; Scott, Colleen N; McCarroll, Matthew E

2013-09-11

364

Efficient production of anti-fluorescein and anti-lysozyme as single-chain anti-body fragments (scFv) by Brevibacillus expression system.  

PubMed

Expression of scFv in Brevibacillus choshinensis was tested using combinations of three different promoters and four different secretion signals. Two model scFv constructs, i.e., His-scFvFLU and His-scFvHEL, were successfully expressed with some of the combinations. Ni Sepharose column and size exclusion chromatography resulted in fairly pure preparations of these two proteins. The purified His-scFvFLU inhibited fluorescence from fluorescein, while the purified His-scFvHEL inhibited lysozyme activity. Relatively high yield of His-scFvFLU (?40%) and His-scFvHEL (?30%) was achieved with the expression and purification system described here. PMID:23973803

Onishi, Hiromasa; Mizukami, Makoto; Hanagata, Hiroshi; Tokunaga, Masao; Arakawa, Tsutomu; Miyauchi, Akira

2013-10-01

365

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

NASA Technical Reports Server (NTRS)

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.

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

2002-01-01

366

Analysis of Formation of Pad Stains in Copper Chemical Mechanical Planarization  

NASA Astrophysics Data System (ADS)

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.

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

2009-12-01

367

In vitro viability estimation and preservation of Cenchrus and Pennisetum pollen (Poaceae:Paniceae)  

E-print Network

mM Ca, 2 mM B, and 10 g L' gum agar appeared acceptable for estimating pollen viability for all taxa. In vitro germination, fluorescein diacetate (FDA), and iodine staining were then evaluated on pollen of birdwoodgrass and pearl millet. In vitro... germination and FDA only were evaluated using pollen of common buffelgrass, T-704 buffelgrass, and P. or/en/a/e. A photographic method and a video method of FDA scoring were used in different experiments. There were few cases Ihat showed a significant...

Hill, Jerry Leon

2012-06-07

368

Karyotype analysis of four jewel-beetle species (Coleoptera, Buprestidae) detected by standard staining, C-banding, AgNOR-banding and CMA3/DAPI staining  

PubMed Central

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

Karagyan, Gayane; Lachowska, Dorota; Kalashian, Mark

2012-01-01

369

Protein Reverse Staining: High-Efficiency Microanalysis of Unmodified Proteins Detected on Electrophoresis Gels  

Microsoft Academic Search

A methodology is presented for efficiently gaining structural information from electrophoresed proteins after on-gel detection by imidazole-sodium dodecyl sulfate-zinc reverse staining. As a consequence of reverse staining, (a) protein bands arise transparent against a deep white-stained background, limits of detection being in the femtomole range; (b) there is no loss of image when the gel is kept in distilled water

C. Fernandezpatron; M. Calero; P. R. Collazo; J. R. Garcia; J. Madrazo; A. Musacchio; F. Soriano; R. Estrada; R. Frank; L. R. Castellanosserra; E. Mendez

1995-01-01

370

Investigation of pad staining and its effect on removal rate in copper chemical mechanical planarization  

Microsoft Academic Search

In copper chemical mechanical planarization process, stains are often generated on the pad surface due to the build-up of polishing by-products. Pad staining is a major concern because it might affect defect, non-uniformity across the wafer, and removal rate variation during polishing. In this study, the characteristics of stains formed on an IC1000 XY grooved pad obtained under various polishing

H. Lee; Y. Zhuang; L. Borucki; S. Joh; F. O'Moore; A. Philipossian

2010-01-01

371

High Dynamic Range Microscopy for Color Selective Virtual De-Staining of Immunocytological Specimens  

Microsoft Academic Search

\\u000a Immunocytochemical markers are increasingly applied for diagnosis of diseases. Usually two or more marker stains are applied\\u000a at once, together with a counterstain for a reliable microscopic investigation of cell specimens. As a preprocessing step\\u000a for the detection of marker-positive cells, other stains should be removed by image processing techniques. This virtual de-staining\\u000a can be achieved by color separation algorithms,

David Friedrich; André Bell; Kraisorn Chaisaowong; Till Braunschweig; Ruth Knüchel-Clarke; Til Aach

372

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

Microsoft Academic Search

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

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

1992-01-01

373

Age estimation of blood stains by hemoglobin derivative determination using reflectance spectroscopy.  

PubMed

Blood stains can be crucial in reconstructing crime events. However, no reliable methods are currently available to establish the age of a blood stain on the crime scene. We show that determining the fractions of three hemoglobin derivatives in a blood stain at various ages enables relating these time varying fractions to the age of the blood stain. Application of light transport theory allows addressing the spectroscopic changes in ageing blood stains to changes in chemical composition, i.e. the transition of oxy-hemoglobin into met-hemoglobin and hemichrome. We have found in 20 blood stains that the chemical composition of the blood stain with age, called hemoglobin reaction kinetics, under controlled circumstances, shows a distinct time-dependent behavior, with a unique combination of the three hemoglobin derivatives at all moments in time. Finally, we employed the hemoglobin reaction kinetics inversely to assess the age of 20 other blood stains studied, again over a time period of 0-60 days. We estimated an age of e.g. 55 days correct within an uncertainty margin of 14 days. In conclusion, we propose that the results obtained under controlled conditions demand further evaluation of their possible value for age determination of blood stains on crime scenes. PMID:20729018

Bremmer, Rolf H; Nadort, Annemarie; van Leeuwen, Ton G; van Gemert, Martin J C; Aalders, Maurice C G

2011-03-20

374

A visible DNA-protein stain: Feulgen-Pararosanilin(SO2) Light Green.  

PubMed

Feulgen-Pararosanilin(SO2) Light Green, a DNA-protein stain, is described that is suited both for visual analysis and quantitative cytochemical measurement. The stain has been applied on cervical cells and quantitative aspects have been studied on chicken erythrocyte and rat liver nuclei. In comparison to single staining, the Feulgen-Pararosanilin(SO2) DNA content of the nuclei remains unaltered after application of the combined staining. The Light Green protein content is reduced considerably, however, dependent on the degree of chromatin condensation in the nucleus. PMID:2422687

Oud, P S; Pahlplatz, M M; Hermkens, H G; Tas, J; James, J; Vooijs, G P

1986-01-01

375

Gallocyanin chromalum as a nuclear stain in cytology. I. A cytophotometric comparison of the Husain-Watts Gallocyanin chromalum staining protocol with the Feulgen procedure.  

PubMed

In the present study, the staining characteristics of the Gallocyanin chromalum technique devised by Husain and Watts are compared with the Feulgen reaction. Liver imprints, blood smears, and cervical smears were fixed in ethanol and stained with either the Husain and Watts Gallocyanin chromalum reagent or the Feulgen-Schiff reagent. The slides were then post-treated with 70% ethanol-HCl pH 1.0, or with phosphotungstic acid for 0.5-30 min. The integrated optical density of cell nuclei was measured with a VIDAS image analyzer. In the material stained with the Husain and Watts procedure, some Gallocyanin chromalum was removed from the nuclei in the early phase (5 min) of all the post-treatment steps, followed by a plateau phase where the integrated optical density remained constant for 30 min. In this phase, the nuclear absorbance was highly reproducible and of the same size regardless of the post-treatment. Both the Husain and Watts procedure and the Feulgen-reaction gave quantitative staining of DNA. The Gallocyanin chromalum stain after Husain and Watts is a quick staining procedure for quantitative evaluation of DNA in cytological material. Proper rinsing of the slides is necessary for a good reproducibility of results. PMID:1723725

Schulte, E K; Lyon, H; Prento, P

1991-05-01

376

Fractional contribution of lung, nasal and gastrointestinal absorption to the systemic level following nose-only aerosol exposure in rats: a case study of 3.7-µm fluorescein aerosols  

Microsoft Academic Search

Because absorption takes place from multiple sites of aerosol deposition, it is generally difficult to interpret systemic levels following nose-only inhalation in laboratory rodents. Therefore, this study attempted to determine the fractional contribution of lung, nasal and gastrointestinal (GI) absorption to the observed systemic level following nose-only aerosol exposure in rats using fluorescein as a model powder solute. Rats were

Masahiro Sakagami; Wataru Kinoshita; Kiyoyuki Sakon; Yuji Makino

2003-01-01

377

Dissociation kinetics of macrocyclic trivalent lanthanide complexes of 1,4,7,10-tetraazacyclododecane-1,7-diacetic acid (DO2A).  

PubMed

The [H(+)]-catalyzed dissociation rate constants of several trivalent lanthanide (Ln) complexes of 1,4,7,10-tetraazacyclododecane-1,7-diacetic acid (LnDO2A(+), Ln = La, Pr, Eu, Er and Lu) have been determined in two pH ranges: 3.73-5.11 and 1.75-2.65 at four different temperatures (19-41.0 °C) in aqueous media at a constant ionic strength of 0.1 mol dm(-3) (LiClO(4)). For the study in the higher pH range, i.e. pH 3.73-5.11, copper(II) ion was used as the scavenger for the free ligand DO2A in acetate/acetic acid buffer medium. The rates of Ln(III) complex dissociation have been found to be independent of [Cu(2+)] and all the Ln(III) complexes studied show [H(+)]-dependence at low acid concentrations but become [H(+)]-independent at high acid concentrations. Influence of the acetate ion content in the buffer on the dissociation rate has also been investigated and all the complexes exhibit a first-order dependence on [Acetate]. The dissociation reactions follow the rate law: k(obs) = k(Ac)[Acetate] + K'k(lim)[H(+)]/(1 + K'[H(+)]) where k(AC) is the dissociation rate constant for the [Acetate]-dependent pathway, k(lim) is the limiting rate constant, and K' is the equilibrium constant for the reaction LnDO2A(+) + H(+) ? LnDO2AH(2+). In the lower pH range, i.e. pH 1.75-2.65, the dye indicator, cresol red, was used to monitor the dissociation rate, and all the Ln(III) complexes also show [H(+)]-dependence dissociation pathways but without the rate saturation observed at higher pH range. The dissociation reactions follow the simple rate law: k(obs) = k(H)[H(+)], where k(H) is the dissociation rate constant for the pathway involving monoprotonated species. The absence of an [H(+)]-independent pathway in both pH ranges indicates that LnDO2A(+) complexes are kinetically rather inert. The obtained k(AC) values follow the order: LaDO2A(+) > PrDO2A(+) > EuDO2A(+) > ErDO2A(+) > LuDO2A(+), whereas the k(lim) and k(H) values follow the order: LaDO2A(+) > PrDO2A(+) > ErDO2A(+) > EuDO2A(+) > LuDO2A(+), mostly consistent with their thermodynamic stability order, i.e. the more thermodynamically stable the more kinetically inert. In both pH ranges, activation parameters, ?H*, ?S* and ?G*, for both acetate-dependent and proton-catalyzed dissociation pathways have been obtained for most of the La(III), Pr(III), Eu(III), Er(III) and Lu(III) complexes, from the temperature dependence measurements of the rate constants in the 19-41 °C range. An isokinetic (linear) relationship is found between ?H* and ?S* values, which supports a common reaction mechanism. PMID:21369608

Lin, Chih-Cheng; Chen, Chia-Ling; Liu, Kuan-Yu; Chang, C Allen

2011-06-21

378

Color Correction of Red Blood Cell Area in H&E Stained Images by Using Multispectral Imaging  

Microsoft Academic Search

The color of stained pathological images varies depending on staining conditions. Since pathologists based their diagnosis on changes in color and morphology of a particular tissue component, it is important that color variations in stained pathological images be corrected so as to obtain a more reliable diagnosis. A color correction for Hematoxylin & Eosin (H&E) stained images was proposed previously.

Tokiya Abe; Hideaki Haneishi; Pinky A. Bautista; Yuri Murakami; Masahiro Yamaguchi; Nagaaki Ohyama; Yukako Yagi

379

Platinum blue as an alternative to uranyl acetate for staining in transmission electron microscopy.  

PubMed

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

Inaga, Sumire; Katsumoto, Tetsuo; Tanaka, Keiichi; Kameie, Toshio; Nakane, Hironobu; Naguro, Tomonori

2007-04-01

380

Immunohistochemical staining patterns of canine eyes affected with chronic superficial keratitis.  

PubMed

Fourteen limbal biopsy specimens from 11 dogs with chronic superficial keratitis (CSK) were examined histologically and immunohistochemically. Ten of the 14 specimens had corneal epithelial hyperplasia and/or atrophy. Eleven of the 14 specimens had thickened epithelial basement membranes. Each specimen had cellular infiltration and lamellar disruption of the stroma. An avidin-biotin immunoperoxidase complex stain was used to detect immunoglobulin (Ig) deposition. Twelve of the 14 specimens stained positive for Ig. The staining pattern was consistent and characterized by diffuse deposition of stain in the superficial conjunctival stroma near the limbus. Four of the 12 Ig-positive specimens also stained positive in the superficial corneal stroma with 1 of these 4 also staining positive along the epithelial cell basement membrane. The diffuse pattern of stain deposition and the absence of staining of specific epithelial structures indicated that CSK is not a classical autoimmune disease similar to any disease in the pemphigus group or similar to systemic lupus erythematosus. Although the results may implicate CSK as an immune-mediated disease, nonspecific factors could not be ruled out. PMID:3767102

Eichenbaum, J D; Lavach, J D; Gould, D H; Severin, G A; Paulsen, M E; Jones, R L

1986-09-01

381

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

Microsoft Academic Search

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.

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

1989-01-01

382

Application of Digital Image Analysis and Flow Cytometry To Enumerate Marine Viruses Stained with SYBR Gold†  

PubMed Central

A novel nucleic acid stain, SYBR Gold, was used to stain marine viral particles in various types of samples. Viral particles stained with SYBR Gold yielded bright and stable fluorescent signals that could be detected by a cooled charge-coupled device camera or by flow cytometry. The fluorescent signal strength of SYBR Gold-stained viruses was about twice that of SYBR Green I-stained viruses. Digital images of SYBR Gold-stained viral particles were processed to enumerate the concentration of viral particles by using digital image analysis software. Estimates of viral concentration based on digitized images were 1.3 times higher than those based on direct counting by epifluorescence microscopy. Direct epifluorescence counts of SYBR Gold-stained viral particles were in turn about 1.34 times higher than those estimated by the transmission electron microscope method. Bacteriophage lysates stained with SYBR Gold formed a distinct population in flow cytometric signatures. Flow cytometric analysis revealed at least four viral subpopulations for a Lake Erie sample and two subpopulations for a Georgia coastal sample. Flow cytometry-based viral counts for various types of samples averaged 1.1 times higher than direct epifluorescence microscopic counts. The potential application of digital image analysis and flow cytometry for rapid and accurate measurement of viral abundance in aquatic environments is discussed. PMID:11157214

Chen, Feng; Lu, Jing-rang; Binder, Brian J.; Liu, Ying-chun; Hodson, Robert E.

2001-01-01

383

Environmentally safe removal\\/disposal of Coomassie Brilliant Blue from gel destain and used gel stain  

Microsoft Academic Search

Gel destaining following Coomassie Brilliant Blue (CBB) staining involves the use of toxic reagents. Here we demonstrate the efficacy of various paper adsorbents in adsorbing CBB. Kimwipes adsorbed the best, followed by Teri towels, multifold towels, and Whatman numbers 1 and 3 filter papers. Three Kimwipes completely adsorbed the dye released from a CBB-stained mini-gel. Nonradioactive destain solution can, therefore,

Yaser Dorri; Biji T. Kurien

2010-01-01

384

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

Microsoft Academic Search

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

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

385

Chest staining variation as a signal of testosterone levels in male Verreaux's Sifaka  

Microsoft Academic Search

Male Verreaux's sifaka (Propithecus verreauxi) exhibit variation in the staining of chest hair in association with the activity of the sternal gland. Scent-marking behavior and social relationships have been shown to vary with the state of chest staining. Research on other mammals suggests that sternal gland activity is modulated by testosterone. The goal of this study was to examine the

Rebecca J. Lewis

2009-01-01

386

Silver staining of proteins in polyacrylamide gels Mireille Chevallet, Sylvie Luche and Thierry Rabilloud*  

E-print Network

Silver staining of proteins in polyacrylamide gels Mireille Chevallet, Sylvie Luche and Thierry author email: thierry.rabilloud@cea.fr Phone +33 438 783 212, Fax : +33 438 789 803 Abstract Silver. The sequential phases of silver staining are protein fixation, then sensitization, then silver impregnation

Paris-Sud XI, Université de

387

Environmentally safe removal/disposal of Coomassie Brilliant Blue from gel destain and used gel stain.  

PubMed

Gel destaining following Coomassie Brilliant Blue (CBB) staining involves the use of toxic reagents. Here we demonstrate the efficacy of various paper adsorbents in adsorbing CBB. Kimwipes adsorbed the best, followed by Teri towels, multifold towels, and Whatman numbers 1 and 3 filter papers. Three Kimwipes completely adsorbed the dye released from a CBB-stained mini-gel. Nonradioactive destain solution can, therefore, be recycled for destaining CBB-stained gels. Stain removal with Kimwipes helps in reducing destain use and in reducing organic liquid waste, and it is 7.5-fold cheaper compared with an available method for CBB disposal. Following this, we determined the suitability of this procedure to remove the dye from a used CBB staining solution awaiting proper disposal by our Institutional Safety Office. The dye from a 0.05% CBB staining solution could be removed in 5 to 10 min using 75 Kimwipes. The CBB-adsorbed Kimwipes did not release the stain when squeezed dry even after incubation in various salts over 1week and in water for 5 weeks. The CBB removed allows its easy disposal as solid waste and will not leach out from solid landfills. Thus, stain removal with Kimwipes helps in disposing CBB in an environmentally friendly manner and allows recycling of destaining solution. PMID:20507825

Dorri, Yaser; Kurien, Biji T

2010-09-15

388

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

Microsoft Academic Search

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

Sheldon Wolff; Paul Perry

1974-01-01

389

Blood-Stained Colostrum and Human Milk during Pregnancy and Early Lactation.  

PubMed

Blood-stained colostrum occurs occasionally during pregnancy and lactation due to a conspicuous increase in lobuloalveolar growth. We report on a case of bilateral frank blood-stained colostrum secreted during pregnancy and early postpartum, emphasizing the transitory nature of this condition and the need to reinforce breastfeeding. PMID:24925862

Barco, Israel; Vidal, M Carmen; Barco, José; Badia, Angels; Piqueras, Mercè; García, Antonio; Pessarrodona, Antoni

2014-11-01

390

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

PubMed

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

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

2012-12-01

391

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

NASA Astrophysics Data System (ADS)

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.

Wang, Gang; Yan, Dongmei; Yang, Yang

2010-10-01

392

Modified detergent Ziehl-Neelsen technique for the staining of Cyclospora cayetanensis.  

PubMed Central

Cyclospora cayetanensis is a cause of prolonged diarrhoea, mainly in travellers. Laboratory diagnosis may be achieved by a number of methods such as the staining of faecal smears by the modified Ziehl-Neelsen (ZN) technique. Safer methods using this technique have been described for the staining of acid fast bacilli and cryptosporidia by replacing the phenol content of the carbol fuschin stain with various concentrated detergents. In this report the technique was modified slightly using a non-concentrated detergent and applied to the staining of oocysts of C cayetanensis. It was found that oocysts of C cayetanensis do not stain using the modified detergent ZN method when compared with similar preparations containing oocysts of Cryptosporidium spp. PMID:8763270

Clarke, S C; McIntyre, M

1996-01-01

393

Characterisation of medieval yellow silver stained glass from Convento de Cristo in Tomar, Portugal  

NASA Astrophysics Data System (ADS)

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.

Delgado, J.; Vilarigues, M.; Ruivo, A.; Corregidor, V.; Silva, R. C. da; Alves, L. C.

2011-10-01

394

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

PubMed

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

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

2014-10-01

395

Yellow staining caused by 4,4'-methylenedianiline exposure. Occurrence among molded plastics workers.  

PubMed

Workers engaged in a molded plastics operation were studied to determine the etiology of yellow staining reactions involving the skin, nails, and hair. A walk-through survey of the facility, medical interviews, physical examinations, and blood and urine tests were performed. 4,4'-Methylenedianiline (MDA), a component chemical of the manufacturing process, produced intense yellow discoloration of nitrocellulose paper in the laboratory and appeared to volatilize readily under ambient conditions. Thirty-five (65%) of 54 process workers showed varying degrees of staining while 11 workers employed in other parts of the factory showed no staining. Yellow staining was restricted to areas of the body where direct contact with MDA appeared likely. Laboratory studies did not provide evidence of systemic toxic effect. Because MDA is a known hepatotoxin for man, with carcinogenic properties in animal test systems, it is important to recognize yellow staining as a cutaneous marker of exposure to this chemical. PMID:4026338

Cohen, S R

1985-08-01

396

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

PubMed Central

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

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

2013-01-01

397

Wet mount microscopy reflects functional vaginal lactobacillary flora better than Gram stain  

PubMed Central

Aim—The status of vaginal lactobacillary flora, an indicator of possible genital infection and pregnancy complications, can be assessed on wet mount or Gram stained specimens. The former is quick, the latter more routine. The accuracy of the two preparative techniques to detect normal vaginal lactobacillary microflora was compared for 646 patients. The effect of delay in transport medium before Gram staining was also investigated. Methods—Patients presented with infectious vaginitis or for a routine prenatal visit. After placement of a speculum, duplicate smears were taken from the upper vaginal vault and examined fresh or after Gram staining. Lactobacillary grades from both methods were compared with lactate concentration in vaginal rinses. In a subgroup of 238 patients, Gram staining was performed both on fresh smears and those that had been transported in Stuart's growth medium. Results—Higher lactobacillary grades (more disrupted flora) were diagnosed 2.9 times more frequently on Gram stained specimens than on wet mounts (p < 0.0001), a difference even more pronounced after transport in Stuart's medium (relative risk, 4.2; p < 0.0001). Lactobacillary grades assessed on wet mounts correlated better with vaginal lactate concentration than those assessed on Gram stains. Conclusions—Easier recognition of lactobacillary morphotypes on wet mounts than on Gram stains might result from the loss of lactobacilli by the process of fixation or Gram staining. Wet mount microscopy of vaginal smears for assessment of lactobacillary grades, rather than Gram staining, is strongly recommended. Key Words: Gram stain • vaginal smears • wet mounts • lactobacillary flora PMID:10823128

Donders, G.; Vereecken, A.; Dekeersmaecker, A.; Van Bulck, B.; Spitz, B.

2000-01-01

398

Transient receptor potential ankyrin 1 activation enhances hapten sensitization in a T-helper type 2-driven fluorescein isothiocyanate-induced contact hypersensitivity mouse model.  

PubMed

Some chemicals contribute to the development of allergies by increasing the immunogenicity of other allergens. We have demonstrated that several phthalate esters, including dibutyl phthalate (DBP), enhance skin sensitization to fluorescein isothiocyanate (FITC) in a mouse contact hypersensitivity model, in which the T-helper type 2 (Th2) response is essential. On the other hand, some phthalate esters were found to activate transient receptor potential ankyrin 1 (TRPA1) cation channels on sensory neurons. We then found a positive correlation between the enhancing effects of several types of phthalate esters on skin sensitization to FITC and their ability to activate TRPA1. Here we examined the involvement of TRPA1 in sensitization to FITC by using TRPA1 agonists other than phthalate esters. During skin sensitization to FITC, the TRPA1 agonists (menthol, carvacrol, cinnamaldehyde and DBP) augmented the ear-swelling response as well as trafficking of FITC-presenting dendritic cells to draining lymph nodes. We confirmed that these TRPA1 agonists induced calcium influx into TRPA1-expressing Chinese hamster ovary (CHO) cells. We also found that TRPA1 antagonist HC-030031 inhibited DBP-induced calcium influx into TRPA1-expressing CHO cells. After pretreatment with this antagonist upon skin sensitization to FITC, the enhancing effect of DBP on sensitization was suppressed. These results suggest that TRPA1 activation will become a useful marker to find chemicals that facilitate sensitization in combination with other immunogenic haptens. PMID:22935519

Shiba, Takahiro; Tamai, Takuma; Sahara, Yurina; Kurohane, Kohta; Watanabe, Tatsuo; Imai, Yasuyuki

2012-11-01

399

Fullerol-fluorescein isothiocyanate-concanavalin agglutinin phosphorescent sensor for the detection of alpha-fetoprotein and forecast of human diseases.  

PubMed

Based on the reaction of the active -OH group in fullerol (F) with the dissociated -COOH group in fluorescein isothiocyanate (FITC) to form an F-FITC and the enhanced effect of N, N-dimethylaniline (DMA) on phosphorescence signal of F-FITC, a new phosphorescent labeling reagent (DMA-F-FITC) was developed. What's more, a phosphorescent sensor for the determination of alpha-fetoprotein variant (AFP-V) has been designed via the coupling technique of the high sensitivity for affinity adsorption-solid substrate-room temperature phosphorimetry (AA-SS-RTP) with the strong specificity reaction between DMA-F-FITC-Con A and AFP-V. The DMA-F-FITC increased the number of luminescent molecules in the biological target which improved the sensitivity of phosphorescent sensor. The proposed sensor was responsive, simple, selective and sensitive, and it has been applied to the determination of trace AFP-V in human serum and the forecast of human diseases using phosphorescence emission wavelength of F or FITC, with the results agreed well with those obtained by enzyme-linked immunoassay (ELISA). Meanwhile, the mechanisms for the labeling reaction and the sensing detection of AFP-V were discussed. PMID:23832221

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

400

Quantitative alpha-ketoglutarate dehydrogenase activity staining in brain sections and in cultured cells.  

PubMed

The activity of a key mitochondrial enzyme, the alpha-ketoglutarate dehydrogenase complex (KGDHC), declines in the brains of patients with neurodegenerative diseases such as Alzheimer's disease, as well as in thiamine-deficient (TD) animals. The decreased activity often occurs without a reduction in enzyme protein, which negates the use of immunocytochemistry to study cellular or regional changes in enzyme activity within the brain. To overcome this limitation, an activity staining method using nitroblue tetrazolium was developed. The histochemical activity staining was standardized in cultured cells. The assay was linear with time and was highly specific for KGDHC. The dark-blue reaction product (formazan) formed a pattern that was consistent with mitochondrial localization. Treatment of the cultured cells with both reversible and irreversible inhibitors decreased formazan production, whereas conventional enzyme assays on cell lysates only revealed loss of KGDHC activity with irreversible inhibitors. The activity staining was also linear with time and highly specific for KGDHC activity in mouse brain sections. Staining occurred throughout the brain, and discrete neuronal populations exhibited particularly intense staining. The pattern of staining differed markedly from the distribution of KGDHC protein by immunocytochemistry. Generalized decreases in the intensity of activity staining that occurred in the TD brains compared to controls were comparable with the loss of KGDHC activity by conventional enzyme assay. Thus, the present study introduces a new histochemical method to measure KGDHC activity at the cellular and regional level, which will be useful to determine changes of in situ enzyme activity. PMID:10610692

Park, L C; Calingasan, N Y; Sheu, K F; Gibson, G E

2000-01-01

401

Acridine orange staining reaction as an index of physiological activity in Escherichia coli  

NASA Technical Reports Server (NTRS)

The assumption that the acridine orange (AO) color reaction may be used as an index of physiological activity was investigated in laboratory grown Escherichia coli. Spectrofluorometric observations of purified nucleic acids, ribosomes and the microscopic color of bacteriophage-infected cells stained with AO confirmed the theory that single-stranded nucleic acids emit orange to red fluorescence while those that are double-stranded fluoresce green in vivo. Bacteria growing actively in a rich medium could be distinguished from cells in stationary phase by the AO reaction. Cells from log phase appeared red, whereas those in stationary phase were green. However, this differentiation was not seen when the bacteria were grown in a minimal medium or when a variation of the staining method was used. Also, shifting bacteria in stationary phase to starvation conditions rapidly changed their AO staining reaction. Boiling and exposure to lethal concentrations of azide and formalin resulted in stationary-phase cells that appeared red after staining but bacteria killed with chlorine remained green. These findings indicate that the AO staining reaction may be suggestive of physiological activity under defined conditions. However, variables in staining and fixation procedures as well as uncertainties associated with mixed bacterial populations in environmental samples may produce results that are not consistent with the classical interpretation of this reaction. The importance of validating the putative physiological implications of this staining reaction is stressed.

McFeters, G. A.; Singh, A.; Byun, S.; Callis, P. R.; Williams, S.

1991-01-01

402

Comparative efficacy of cedarwood oil and xylene in hematoxylin and eosin staining procedures: An experimental study  

PubMed Central

Background: Xylene is used as a clearing agent in hematoxylin and eosin (H and E) staining of tissue sections in routine histopathology based diagnosis. However, the hazards associated with exposure to xylene are of concern. Numerous solutions mainly essential oils have been evaluated in the past as clearing agents, which can possibly be substituted for xylene during the routine tissue processing. Aim: The aim of this study is to compare the efficacy of essential oil (cedarwood oil), as a possible replacement for xylene in H and E staining procedures. Materials and Methods: The study was carried out in the Department of Oral Pathology and Microbiology. Thirty paraffin blocks of the routine biopsy specimen were retrieved from the department archives. The cedarwood oil was procured from organic and essential oil dealer in the local market. Two to three paraffin sections of four micron thickness were cut from each of the 30 paraffin blocks of processed tissue specimens, were subjected to different clearing agents: Essential oil (8% cedarwood oil) or xylene and stained with H and E stain. The stained sections were scored based on nuclear and cytoplasmic details, clarity and uniformity of staining. Results: Significant correlation was observed between cedarwood oil and xylene in terms of the three staining quality parameters assessed. Conclusions: We conclude that cedarwood oil can be an effective, eco-friendly and safe alternative to xylene as a clearing agent in the histopathological laboratory. PMID:25097399

Indu, Sudip; Ramesh, V.; Indu, Priyanka Chakravarty; Prashad, Karthikshree V.; Premalatha, B.; Ramadoss, K.

2014-01-01

403

Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition  

PubMed Central

Single particle electron microscopy (EM), of both negative stained or frozen hydrated biological samples, has become a versatile tool in structural biology 1. In recent years, this method has achieved great success in studying structures of proteins and macromolecular complexes 2, 3. Compared with electron cryomicroscopy (cryoEM), in which frozen hydrated protein samples are embedded in a thin layer of vitreous ice 4, negative staining is a simpler sample preparation method in which protein samples are embedded in a thin layer of dried heavy metal salt to increase specimen contrast 5. The enhanced contrast of negative stain EM allows examination of relatively small biological samples. In addition to determining three-dimensional (3D) structure of purified proteins or protein complexes 6, this method can be used for much broader purposes. For example, negative stain EM can be easily used to visualize purified protein samples, obtaining information such as homogeneity/heterogeneity of the sample, formation of protein complexes or large assemblies, or simply to evaluate the quality of a protein preparation. In this video article, we present a complete protocol for using an EM to observe negatively stained protein sample, from preparing carbon coated grids for negative stain EM to acquiring images of negatively stained sample in an electron microscope operated at 120kV accelerating voltage. These protocols have been used in our laboratory routinely and can be easily followed by novice users. PMID:22215030

Booth, David S.; Avila-Sakar, Agustin; Cheng, Yifan

2011-01-01

404

Identification and age estimation of blood stains on colored backgrounds by near infrared spectroscopy.  

PubMed

Non-destructive identification and subsequent age estimation of blood stains are significant steps in forensic casework. The latter can provide important information on the temporal aspects of a crime. As previously shown, visible spectroscopy of blood stains on white backgrounds can successfully be used for their identification and age estimation. The use of this technique however, is hampered by dark backgrounds. In the present study the feasibility to use near infrared (NIR) spectroscopy was evaluated for blood stain identification and age estimation on dark backgrounds. Using NIR reflectance spectroscopy, blood stains were distinguished from other substances with 100% sensitivity and 100% specificity. In addition, Partial Least Squares Regression analysis was applied to estimate the age of blood stains on colored backgrounds. The age of blood stains up to 1 month old was estimated successfully with a root mean squared error of prediction of 8.9%. These findings are an important step toward the practical implementation of blood stain identification and age estimation in forensic casework, where a large variety of backgrounds can be encountered. PMID:22503886

Edelman, Gerda; Manti, Vicky; van Ruth, Saskia M; van Leeuwen, Ton; Aalders, Maurice

2012-07-10

405

Improved Coomassie Blue Dye-Based Fast Staining Protocol for Proteins Separated by SDS-PAGE  

PubMed Central

The time required to visualize proteins using Coomassie Blue dye has been significantly reduced with the introduction of fast staining protocols based on staining with a Coomassie Blue dye solution at boiling temperatures. However, fast stainings suffer from high gel backgrounds, reducing the signal-to-noise ratio and limiting the number of detectable spots in the case of 2D SDS-PAGE. The aim of this work was to eliminate the high gel background, and thus improve fast staining protocols based on Coomassie Blue dye. We show that merely replacing water with a 4 mM EDTA washing solution at boiling temperatures, results in a transparent gel background within 50 to 60 minutes of destaining. Moreover, when a combination of imidazole-zinc reverse staining and Coomassie Blue-based fast staining is used the sensitivity is improved significantly; nanogram amounts of proteins can be detected using 1D SDS-PAGE, and about 30% to 60% more spots can be detected with 2D SDS-PAGE in plasma, platelet, and rat brain tissue samples. This work represents an optimized fast staining protocol with improved sensitivity, requiring between 60 to 75 minutes to complete protein visualization. PMID:24278455

Majek, Pavel; Riedelova-Reicheltova, Zuzana; Pecankova, Klara; Dyr, Jan E.

2013-01-01

406

Differential dichrome staining of tissue culture monolayers: alternate dyes and possible mechanism.  

PubMed

A polyacid-dependent dichrome has been devised which will differentiate epithelial from mesenchymal cells in young dividing primary cultures. Epithelial cells and colonies and nuclei are stained with metanil yellow, the stain is fixed and differentiated with phosphotungstic acid, and the mesenchymal elements are stained with toluidine blue. Several other dyes are tested for substitution in this method. Biebrich scarlet and aniline blue could be substituted for the metanil yellow; Bismarck brown T, Janus green B, crystal violet, and neutral red could be substituted for the basic dye. PMID:89718

Everett, M M; Miller, W A

1978-11-01

407

New tetrachromic VOF stain (Type III-G.S) for normal and pathological fish tissues.  

PubMed

A new VOF Type III-G.S stain was applied to histological sections of different organs and tissues of healthy and pathological larvae, juvenile and adult fish species (Solea senegalensis; Sparus aurata; Diplodus sargo; Pagrus auriga; Argyrosomus regius and Halobatrachus didactylus). In comparison to the original Gutiérrez VOF stain, more acid dyes of contrasting colours and polychromatic/metachromatic properties were incorporated as essential constituents of the tetrachromic VOF stain. This facilitates the selective staining of different basic tissues and improves the morphological analysis of histochemical approaches of the cell components. The VOF Type III -6.5 stain is composed of a mixture of several dyes of varying size and molecular weight (Orange Gstained. Muscle fibers, collagen, reticulin and elastin fibers, erythrocytes, cartilage, bone, mucous cells, oocytes and larvae were selectively stained and differentiated. Dyes with small size and molecular weight (i.e Orange G), penetrate all tissue structures rapidly, but are only tightly retained in densely textured tissues (i.e erythrocytes). Methyl Blue is an interesting triarylmethane dye (large size and molecular weight), which is incorporated in this new VOF tetrachrome stain, and acquires histochemical significance when used at acid pH (2.8) because collagen and reticulin fibers, as well basophilic and metachromatic substances (strongly ionized sulphated glycoconjugates) can be identified. Muscle tissues show an evident green colour (Fast Green or Light Green affinities), even those isolated and/or diffuse muscle fibers present in the digestive submucosa layer. Connective tissues showed a specific and strong blue colour (Methyl Blue affinity) or mixed blue-red staining (Methyl Blue and Acid Fucshin affinities). Very noticeable is the staining of the mucous cells, as well as the hyaline capsule of the viral lymphocystic cells, which were stained blue-purple (carboxylated and/or strongly ionized sulphated groups). Cartilaginous tissues showed a blue or purple (Methyl Blue affinity) staining, and a specific red colour (Acid Fucshin affinity) was evident during calcification or in bone structures (i.e skeleton, fins, gills, teeth). PMID:15967749

Sarasquete, C; Gutiérrez, M

2005-01-01

408

A rapid method for preparing high quality alizarin stained skeletons of adult mice.  

PubMed

A simple three-day technique is described for preparing completely cleared and high quality alizarin stained total skeletons of adult mice. Unfixed specimens are partially macerated during staining. Older specimens are heated for 15 min in 1% KOH. A heated solution of benzyl and ethyl alcohol, glycerin, and water is used for final clearing and hardening. This procedure requires about 10 min work per specimen and greatly simplifies preparation of stained and cleared skeletons of adult mice. Another technique, giving slightly better preparations, but requiring 11-14 days, is also described. PMID:2441491

Selby, P B

1987-05-01

409

Development of a Whole Blood Staining Device for use During Space Shuttle Flights  

NASA Technical Reports Server (NTRS)

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.

Sams, Clarence F.; Crucian, Brian E.; Clift, Vaughan L.; Meinelt, Ellen M.

1999-01-01

410

Topical dual-stain difference imaging for rapid intra-operative tumor identification in fresh specimens  

PubMed Central

Assessing tumor margin status during surgery is critical to ensure complete resection of cancer tissue; however, current approaches are ineffective and often result in repeat surgery. We present a novel optical imaging approach for margin assessment using topical application of two fluorescent stains, one targeted to a tumor biomarker and the other a non-targeted reference, to freshly excised specimens. Computing a normalized difference image from fluorescence images of the targeted and untargeted stains suppresses the confounding effects of non-specific uptake. Applying this approach in excised breast tumor models produced promising tumor-to-normal tissue contrasts that were significantly higher than single-targeted-stain imaging. PMID:24281541

Davis, Scott C.; Gibbs, Summer L.; Gunn, Jason R.; Pogue, Brian W.

2014-01-01

411

Application of Prussian blue staining in the diagnosis of ocular siderosis  

PubMed Central

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

Yang, Zhen; Yang, Xiao-Li; Xu, Li-Shuai; Dai, Le; Yi, Mei-Chao

2014-01-01

412

Under-air staining of the anterior capsule using Trypan blue with a 30 G needle  

PubMed Central

The original technique of staining the anterior capsule of the lens with Trypan blue involves the injection of an air bubble in the anterior chamber. A drawback of this technique is the possible instability of the anterior chamber caused by the sudden exit of air when the dye is injected with the cannula through the side-port incision. Other staining techniques that use viscoelastic substances to increase the stability of the anterior chamber and to dose the injected dye have been described. The authors present an under-air staining technique of the anterior capsule using one drop of Trypan blue injected with a 30 G needle through the peripheral cornea. This procedure prevents the air bubble from escaping the anterior chamber and allows fast and selective staining of the capsule. PMID:23386783

Giammaria, Daniele; Giannotti, Michele; Scopelliti, Angelo; Pellegrini, Giacomo; Giannotti, Bruno

2013-01-01

413

Phosphoprotein staining for sodium dodecyl sulfate-polyacrylamide gel electrophoresis using fluorescent reagent morin hydrate.  

PubMed

A fluorescence-based stain with 3,5,7,2',4'-pentahydroxyflavone (morin hydrate, MH) was designed to stain phosphoproteins in one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Al(3+) was applied as a "fixed bridge," providing an efficient energy transfer channel between phosphoprotein and MH, to produce a strong fluorescent complex for the determination of phosphoprotein. As little as 62.5ng of ?-casein (7 or 8 phosphates) and ?-casein (5 phosphates), 125ng of ovalbumin (2 phosphates), and ?-casein (1 phosphate) could be visualized with a wide linear dynamic range. In comparison with conventional methods, MH stain is a time-saving method that takes just 90min. It also has good compatibility with routine protein stainings such as Coomassie Brilliant Blue R (CBBR) and SYPRO Ruby for total protein analysis. PMID:23274386

Wang, Xu; Hwang, Sun-Young; Cong, Wei-Tao; Jin, Li-Tai; Choi, Jung-Kap

2013-04-01

414

The staining of starch gels with Coomassie Brilliant Blue G 250 perchloric acid solution  

Microsoft Academic Search

Summary Coomassie Brilliant Blue G250 dissolved in dilute perchloric acid solution was used to stain protein bands in starch gels. The bands were visible within min, no destaining was required and the gels could be stored indefinitely.

Gail McFarland

1977-01-01

415

A new method of urinary stone analysis by batik histochemical staining of thin sections.  

PubMed

Thin sections of urinary calculi are prepared by petrographic methods using Araldite as the mounting medium. By covering the remaining part of the section with wax, an exposed segment of the section is stained by a histochemical technique. By the process of dewaxing and rewaxing, successive adjacent segments are stained by GBHA, Von Kossa, Schultz, and titan yellow methods for calcium oxalate, apatite, uric acid and urates, and magnesium in magnesium ammonium phosphate, respectively. If desired, matrix in additional segments is stained with PAS and aqueous toluidine blue. Microscopic examination of each layer through all the stained segments of a stone section reveals its chemical nature. Thus the chemical composition, morphology, and spatial distribution of the crystalline and matrix constituents of thin sections of urinary calculi are simultaneously revealed in situ. PMID:91241

Kabra, V; Kabra, S G

1979-05-01

416

DNA the Easy Way (And âÂÂGram Stainâ Without the Mess)  

NSDL National Science Digital Library

This resource is a short laboratory exercise that teaches students the procedures of DNA isolation from bacterial cells. In addition, students learn how to determine the Gram-stain reaction of bacterial isolates.

Gail L. Schumann (University of Massachusetts;); Claudia A. Jasalavich (Nashua, NH;)

2001-02-22

417

Characterization of organic emission from a wood finishing product-wood stain  

SciTech Connect

The paper gives results of the measurement of emission characteristics of four organic compounds (nonane, decane, undecane, and 1,2,4-trimethylbenzene) from a wood finishing product, wood stain, in an environmental chamber. It was found that the emission patterns of the four organic compounds can be described by a two-phase model: phase 1, when the wood stain is relatively wet; and phase 2, when the wood stain becomes relatively dry. The changes of emission mechanisms between phases 1 and 2 were reflected by the significantly different emission and decay rates measured during the two periods. A relationship was found that can be used to predict the relative emission and decay rates of the four organic compounds from the wood stain.

Chang, J.C.S.; Guo, Z.

1992-01-01

418

Prospective study of gram-stained stool smears in diagnosis of Clostridium difficile colitis.  

PubMed Central

Gram stains of stools from patients with diarrhea and control patients with no diarrhea were examined for a predominance of gram-positive rods and the presence of polymorphonuclear leukocytes. Results were compared with those from lower gastrointestinal endoscopy for pseudomembranes. Clostridium difficile culturing, and C. difficile toxin assay. The Gram stain was moderately difficult to interpret and was not useful in diagnosing diarrheal disease associated with C. difficile. PMID:6190839

Shanholtzer, C J; Peterson, L R; Olson, M N; Gerding, D N

1983-01-01

419

Weathering patinas on the medieval (S. XIV) stained glass windows of the Pedralbes Monastery (Barcelona, Spain)  

Microsoft Academic Search

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

Meritxell Aulinas; Maite Garcia-Valles; Domingo Gimeno; Jose Luis Fernandez-Turiel; Flavia Ruggieri; Montserrat Pugès

2009-01-01

420

Study of stain-eliminating textiles using ZnO nanoparticles  

Microsoft Academic Search

We report the synthesis and characterization of nano-sized zinc oxide (ZnO) particles and their application on cotton and polyester\\/cotton (P\\/C) fabrics for imparting and evaluating the stain-eliminating or stain-release function by surface modification. The ZnO nanoparticles were produced in different conditions of temperature (90°C or 150°C) and reacting medium (water or 1,2-ethanediol). A high temperature was necessary to obtain small

S. Kathirvelu; Louis DSouza; Bhaarathi Dhurai

2010-01-01

421

Vasectomy with rivanol injection and fertility control by vital staining with eosin.  

PubMed

A material is presented of 66 males who were treated with Rivanol injection during vasectomy in order to obtain immediate sterility. The fertility control was established using vital eosin staining on seminal fluid at home. One case of recanalization occurred. All patients had infertile spermatozoa or aspermia 10 days after surgery. Increased frequency of complications was not observed. Vital staining with eosin is found to be practical and easily used. PMID:2448262

Lauritsen, N P; Kløve-Mogensen, M; Glavind, K

1987-01-01

422

P16, EGFR, Cyclin D1, and p53 Staining Patterns for Inverted Papilloma  

PubMed Central

Introduction We aim to better characterize the staining patterns of inverted papilloma (IP) with and without carcinoma by performing immunohistochemistry for p16, EGFR (Epidermal Growth Factor Receptor), p53, and Cyclin D1 antibodies on a large patient cohort. Methods One hundred and sixty-two IP specimens from 122 patients treated at the University of Michigan between 1996 and 2011. Twenty-two specimens contained carcinoma. Tumor was extracted for construction of two tissue microarrays and stained for p16, EGFR, p53, and Cyclin D1. Tumor staining intensity and percentage staining were scored. Results Mean percentage staining for IP and IP with carcinoma was 12% versus 7% for p16 (no statistical significance, NS), 20% versus 34% for EGFR (NS), 4% versus 24% for p53 (p<0.001), and 17% versus 21% for Cyclin D1 (NS). Benign disease was positive for p16 in 64%, EGFR in 50%, p53 in 30%, and Cyclin D1 in 76%. Inverted papilloma with carcinomatous degeneration was positive for p16 in 14%, EGFR in 71%, p53 in 62%, and Cyclin D1 in 76%. This is statistically significant for differences between IP and IP carcinoma for p16 and p53 staining only. Conclusion Important characteristic staining pattern for inverted papilloma with and without carcinoma are highlighted in this study. Unlike recent trends in HPV-related head and neck malignancies, low expression of p16 is a marker for malignancy in this series. Positive staining for p53 correlates with the development of carcinoma in inverted papilloma. PMID:24039221

Lin, Giant C.; Scheel, Adam; Akkina, Sarah; Chinn, Steven; Graham, Martin; Komarck, Christine; Walline, Heather; McHugh, Jonathan B.; Prince, Mark E.; Carey, Thomas; Zacharek, Mark A.

2014-01-01

423

A new panoptic stain for developmental biology--the mouse placenta paradigm.  

PubMed

A panoptic histological stain, PHTA, is introduced for routine use in developmental biology. The protocol is based on three dyes, hematoxylin and, after tannic acid treatment, phloxine B and azure B. It was optimized for differential staining of extracellular matrix, cytoplasm and chromatin. The method is quick, inexpensive and well-reproducible. The mouse placenta is used here to demonstrate the excellent suitability of PHTA for routine morphology. PMID:11677809

Kurz, H; Wittekind, D

2001-09-01

424

Detection of alkali-silica reaction swelling in concrete by staining  

DOEpatents

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.

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

1998-01-01

425

Detection of alkali-silica reaction swelling in concrete by staining  

DOEpatents

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.

Guthrie, G.D. Jr.; Carey, J.W.

1998-04-14

426

Immunoperoxidase staining for identification of Aspergillus species in routinely processed tissue sections.  

PubMed Central

AIMS: To evaluate the performance of an immunoperoxidase stain using the monoclonal antibody EB-A1 to detect Aspergillus species in formalin fixed, paraffin wax embedded tissue. METHODS: The monoclonal antibody EB-A1 directed against galactomannan was used to detect Aspergillus species in 23 patients with suspected or confirmed invasive aspergillosis. Immunostaining was performed on formalin fixed, paraffin wax embedded tissue using the streptavidin-biotin method and compared with conventional haematoxylin and eosin, periodic acid-Schiff, and Gomori-Grocott stains. Results of immunostaining were semiquantitatively analysed with regard to both intensity of staining and number of positively staining micro-organisms. Tissue sections from 16 patients with confirmed invasive mycoses due to Candida species, Apophysomyces elegans, Rhizopus oryzae, Pseudallescheria boydii and Histoplasma capsulatum were used as controls. RESULTS: In 19 (83%) of 23 cases invasive aspergillosis was confirmed by both histological examination and culture (18 Aspergillus fumigatus and one A flavus). Immunoperoxidase stains were positive in 17 (89%) of 19 cases including one case of disseminated infection due to A flavus. Furthermore, the immunoperoxidase stain was positive in a culture negative tissue section with histological evidence of mycelial development, indicating the presence of Aspergillus species. Some cross-reactivity was observed with the highly related fungus P boydii, although the number of mycelial elements that stained was low. CONCLUSIONS: Immunoperoxidase staining using the monoclonal antibody EB-A1 performs well on routinely processed tissue sections and permits detection and generic identification of Aspergillus species, although it was no better than conventional histopathology in identifying the presence of an infection. An additional advantage is that the immunostain may help to provide an aetiological diagnosis when cultures remain negative. Images PMID:8943743

Verweij, P E; Smedts, F; Poot, T; Bult, P; Hoogkamp-Korstanje, J A; Meis, J F

1996-01-01

427

Comparison of Chemicon SimulFluor Direct Fluorescent Antibody Staining with Cell Culture and Shell Vial Direct Immunoperoxidase Staining for Detection of Herpes Simplex Virus and with Cytospin Direct Immunofluorescence Staining for Detection of Varicella-Zoster Virus  

Microsoft Academic Search

A new rapid direct immunofluorescence assay, the SimulFluor direct fluorescent-antibody (DFA) assay, which can simultaneously detect herpes simplex virus types 1 and 2 (HSV-1 and -2) and varicella-zoster virus (VZV), was evaluated in comparison with our current standard procedures of (i) shell vial direct immuno- peroxidase (shell vial IP) staining and cell culture for detection of HSV and (ii) cytospin

EDWARD L. CHAN; KEN BRANDT; GREG B. HORSMAN

2001-01-01

428

Transient receptor potential ankyrin 1 activation enhances hapten sensitization in a T-helper type 2-driven fluorescein isothiocyanate-induced contact hypersensitivity mouse model  

SciTech Connect

Some chemicals contribute to the development of allergies by increasing the immunogenicity of other allergens. We have demonstrated that several phthalate esters, including dibutyl phthalate (DBP), enhance skin sensitization to fluorescein isothiocyanate (FITC) in a mouse contact hypersensitivity model, in which the T-helper type 2 (Th2) response is essential. On the other hand, some phthalate esters were found to activate transient receptor potential ankyrin 1 (TRPA1) cation channels on sensory neurons. We then found a positive correlation between the enhancing effects of several types of phthalate esters on skin sensitization to FITC and their ability to activate TRPA1. Here we examined the involvement of TRPA1 in sensitization to FITC by using TRPA1 agonists other than phthalate esters. During skin sensitization to FITC, the TRPA1 agonists (menthol, carvacrol, cinnamaldehyde and DBP) augmented the ear-swelling response as well as trafficking of FITC-presenting dendritic cells to draining lymph nodes. We confirmed that these TRPA1 agonists induced calcium influx into TRPA1-expressing Chinese hamster ovary (CHO) cells. We also found that TRPA1 antagonist HC-030031 inhibited DBP-induced calcium influx into TRPA1-expressing CHO cells. After pretreatment with this antagonist upon skin sensitization to FITC, the enhancing effect of DBP on sensitization was suppressed. These results suggest that TRPA1 activation will become a useful marker to find chemicals that facilitate sensitization in combination with other immunogenic haptens. -- Highlights: ? Role of TRPA1 activation was revealed in a mouse model of skin sensitization to FITC. ? TRPA1 agonists enhanced skin sensitization as well as dendritic cell trafficking. ? Dibutyl phthalate (DBP) has been shown to enhance skin sensitization to FITC. ? TRPA1 activation by DBP was inhibited by a selective antagonist, HC-030031. ? HC-030031 inhibited the enhancing effect of DBP on skin sensitization to FITC.

Shiba, Takahiro; Tamai, Takuma; Sahara, Yurina; Kurohane, Kohta [Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, 52?1 Yada, Suruga-ku, Shizuoka City, Shizuoka 422?8526 (Japan)] [Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, 52?1 Yada, Suruga-ku, Shizuoka City, Shizuoka 422?8526 (Japan); Watanabe, Tatsuo [Laboratory of Food Chemistry, School of Food and Nutritional Sciences, University of Shizuoka, 52?1 Yada, Suruga-ku, Shizuoka City, Shizuoka 422?8526 (Japan)] [Laboratory of Food Chemistry, School of Food and Nutritional Sciences, University of Shizuoka, 52?1 Yada, Suruga-ku, Shizuoka City, Shizuoka 422?8526 (Japan); Imai, Yasuyuki, E-mail: imai@u-shizuoka-ken.ac.jp [Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, 52?1 Yada, Suruga-ku, Shizuoka City, Shizuoka 422?8526 (Japan)] [Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, 52?1 Yada, Suruga-ku, Shizuoka City, Shizuoka 422?8526 (Japan)

2012-11-01

429

Effects of short-term portacaval anastomosis on the peripheral and brain disposition of the blood-brain barrier permeability marker sodium fluorescein in rats.  

PubMed

Contradictory results have been reported with regard to the effects of various models of hepatic encephalopathy on the blood-brain barrier (BBB) permeability, which may be due partly to the use of brain concentrations of BBB markers without attention to their peripheral pharmacokinetics. The purpose of the current study was to investigate the effects of short-term portacaval anastomosis (PCA), a type B model of hepatic encephalopathy, on the peripheral pharmacokinetics and brain distribution of sodium fluorescein (FL), which is a small molecule marker of BBB passive permeability. A single 25mg/kg dose of FL was administered intravenously to 10-day PCA and sham-operated rats, and serial blood and bile (0-30min) and terminal (30min) brain samples were collected, and the concentrations of FL and its glucuronidated metabolite (FL-Glu) were measured by HPLC. Additionally, the free fractions of FL (fu) in all the plasma samples were determined, and the effects of bile salts on fu were investigated in vitro. Passive permeability of BBB to FL was estimated by brain uptake clearance (Kin) based on both the brain concentrations of FL and plasma concentrations of free (unbound) FL. PCA caused a 26% increase in the fu of FL in plasma, which was due to competition of bile acids with FL for binding to plasma proteins. Additionally, PCA reduced the biliary excretion of FL-Glu by 55%. However, free Kin values (µl/min/g brain) for the sham (0.265±0.034) and PCA (0.228±0.038) rats were not significantly different. It is concluded that whereas 10-day PCA alters the peripheral pharmacokinetics of FL, it does not significantly affect the BBB permeability to the marker. PMID:23916670

Shaik, Imam H; Miah, Mohammad K; Bickel, Ulrich; Mehvar, Reza

2013-09-19

430

Single-step procedure for labeling DNA strand breaks with fluorescein- or BODIPY-conjugated deoxynucleotides: Detection of apoptosis and bromodeoxyuridine incorporation  

SciTech Connect

The methods of in situ labeling of DNA strand breaks have been used to identify apoptotic cells and/or DNA replicating cells. While discrimination of apoptotic cells is based on the inherent presence of numerous DNA strand breaks in their chromatin, DNA proliferating cells can be discriminated by the selective DNA strand break induction by photolysis (SBIP) methodology at the sites that contain incorporated bromodeoxyuridine (BrdUrd) or iododeoxyuridine (IdUrd). In both instances, DNA strand breaks are labeled with biotin- or digoxygenin-conjugated deoxynucleotides, preferably in the reaction catalyzed by exogenous terminal deoxynucleotidyl transferase; fluorescein tagged avidin (streptavidin) or digoxygenin antibody is used in the second step of the reaction. In the present study, DNA strand break labeling was simplified by using directly labeled deoxynucleotides, in a single-step reaction. Apoptotic cells in HL-60 cultures treated with camptothecin or in primary cultures of non-Hodgkin`s lymphoma cells treated with prednisolone were easily identified utilizing BODIPY-conjugated dUTP (B-dUTP). The single-step procedure, requiring fewer centrifugation steps, resulted in less cell loss compared to the two-step cell labeling technique. The morphology of cells subjected to SBIP was excellent, allowing visualization of distinct DNA replication points. Because, unlike the immunocytochemical methods used to detect BrdUrd incorporation, the SBIP methodology does not require DNA denaturation by heat or acid, nuclear proteins are expected to remain undenatured in situ, allowing one to study colocalization of various constituents, detected immunocytochemically, at the DNA replication points. 30 refs., 7 figs.

Xun Li; Traganos, F.; Melamed, M.R.; Darzynkiewicz, Z. [New York Medical College, Valhalla, NY (United States)

1995-06-01

431

Modification of Luna's technique for staining eosinophils in the hamster cheek pouch.  

PubMed

The cheek pouch is an anatomical peculiarity of hamsters, widely used as an experimental model for oral cancer, and characterization of its normal cell populations and the changes they undergo in pathological conditions is of great interest. Our studies of epithelium-connective tissue interactions have revealed that hamster eosinophils are not easily recognizable because they are small and exhibit a larger nucleus:cytoplasm ratio than those in human and other animal tissues. Luna's technique is the most popular specific staining technique for eosinophils. Owing to the morphology of hamster eosinophils, however, it was necessary to modify Luna's technique to stain these cells selectively against a more contrasting background that would enable their identification and quantitation in the hamster cheek pouch. The modification involved staining the sections with a solution of 0.5% Biebrich scarlet in lithium carbonate followed by counterstaining with 1% metanil yellow in water. The eosinophils were stained selectively red against a yellow background. Our technique avoided nuclear staining and enhanced observation of selectively stained granules in a scarce cytoplasm with a contrasting background, which permits fast, reproducible studies and automated image analysis. PMID:18802813

Tomasi, V H; Pérez, M A; Itoiz, M E

2008-06-01

432

Selective axonal argentaffin staining in rat central nervous system after protein mercuration.  

PubMed

The mercury-silver (Hg-Ag) argentaffin technique, known to stain specifically proteins in the lateral components of triads/diads in striated muscle cells, was applied to the central nervous system of adult rats. Following fixation in glutaraldehyde, axons in white and gray matter were selectively stained, but not perikarya or their proximal axon and dendrites. Neural tissues were postfixed 24 hr in 5% (w/v) mercuric acetate in 2% (v/v) acetic acid in distilled water, stained for 12-24 hr in darkness at 37-43 C with ammoniacal silver nitrate solution, freshly prepared by adding concentrated ammonia to 60% (w/v) silver nitrate solution until a small amount of silver oxide precipitate remained undissolved. Samples were then washed with freshly prepared 5% (w/v) sodium sulfite and distilled water. All steps were carried out using dark-colored glass flasks. Samples were dehydrated with ethanol and embedded in Paraplast or Poly Bed. Electron microscopy showed the silver-reducing protein inside the axons. Methylation abolished Hg-Ag axonal reactivity indicating that carboxyl groups were necessary for silver staining. Proteins with solubility properties characteristic of neurofilament proteins were involved in Hg-Ag staining. In the cerebellum the plexus of parallel fibers in the molecular layer were not stained, while basket cell axonal processes reacted intensely. The method appears to distinguish neuronal protein variants related to cytotypic differences in cytoskeletal neurofilaments. PMID:7703304

Tandler, C J; Pellegrino de Iraldi, A

1994-11-01

433

Staining human lymphocytes and onion root cell nuclei with madder root.  

PubMed

We performed staining experiments on cells using natural dyes and different mordants using techniques that are used for wool and silk dyeing. The natural dye sources were madder root, daisy, corn cockle and yellow weed. Ferrous sulfate, copper sulfate, potassium tartrate, urea, potassium aluminum sulfate and potassium dichromate were used as mordants. Distilled water, distilled water plus ethanol, heptane, and distilled water plus methanol were used as solvents. All dye-mordant-solvent combinations were studied at pH 2.4, 3.2 and 4.2. The generic staining procedure was to boil 5-10 onion roots or stimulated human lymphocyte (SHL) preparations in a dye bath on a hot plate. Cells were examined at every half hour. For multicolor staining, madder-dyed lymphocytes were decolorized, then stained with Giemsa. The AgNOR technique was performed following the decolorization of Giemsa stained lymphocytes. Good results were obtained for both onion root cells and lymphocytes that were boiled for 3 h in a dye bath that included 4 g madder root, 4 g ferrous sulfate as mordant in 50 ml of 1:1 (v/v) methanol:distilled water. The pH was adjusted to 4.2 with 6 ml acetic acid. We conclude that madder root has potential as an alternative dye for staining biological materials. PMID:15804822

Cücer, N; Guler, N; Demirtas, H; Imamo?lu, N

2005-01-01

434

Standardization of the Feulgen-Schiff technique. Staining characteristics of pure fuchsin dyes; a cytophotometric investigation.  

PubMed

Four fuchsin analogues (Pararosaniline, Rosaniline. Magenta II and New Fuchsin) usually found in Basic Fuchsin have been applied as chemically pure dyes to the Feulgen-technique. Total nuclear absorption and wavelength of the absorption maximum were measured by microspectrophotometry in Feulgen stained cytological and plastic embedded histological liver samples, and in lymphocyte nuclei in human peripheral blood smears; absorption spectra of Feulgen stained DNA-polyacrylamide films were determined by spectrophotometry. The grey value distribution of tetraploid liver cell nuclei was calculated with an image analyzer. The staining characteristics of the pure dyes were compared to commercial fuchsin samples from various suppliers. Reverse phase thin layer chromatography was used for characterization and qualitative separation of commercial batches. Pure fuchsin analogues were all equally suitable for Feulgen staining: with respect of staining intensity all pure fuchsin dyes gave nearly identical results with a bathochromic shift of the absorption maximum from Pararosaniline to New Fuchsin of about 8 microns. Differences in staining results observed among the commercial dyes were due to varying dye content, contamination with an acridine-like fluorescent compound or simply mislabelling of samples. Pure Pararosaniline is recommended for a standard Feulgen technique. PMID:2732097

Schulte, E; Wittekind, D

1989-01-01

435

Journal of Neuroscience Methods 153 (2006) 135146 A modified technique for high-resolution staining of myelin  

E-print Network

of colloidal silver to myelin for vi% formalin following impregnation in ammoniacal silver nitrate, and the use of a low concentration of 4 reveals similar patterns in the new myelin silver stain, the Gallyas stain, and myelin basic protein

Wang, Xiaoqin

436

A rapid method of staining ultrathin sections for surgical pathology TEM with the use of the microwave oven.  

PubMed

A rapid microwave method is described for staining ultrathin sections for surgical pathology transmission electron microscopy (TEM). Three sets of Epon sections of human heart biopsy were mounted on unsupported 200-mesh Rhodium-coated copper grids and were stained with uranyl acetate (UA) and lead citrate. The first set of grids was stained for 15 seconds in each solution with the aid of a microwave oven, and the second set was stained routinely for 30 minutes in UA and 10 minutes in lead citrate at room temperature. The third control set was stained for 15 seconds in each solution without microwave bombardment. The overall image quality of the TEM micrographs generated by the "quick-stained" microwave enhanced sections was better than routine stained and control sections. The microwave-treated sections have more contrast, less artifacts in the form of precipitate, and a more uniform overall staining. PMID:2581442

Estrada, J C; Brinn, N T; Bossen, E H

1985-05-01

437

Identification of gentian violet concentration that does not stain oral mucosa, possesses anti-candidal activity and is well tolerated  

Microsoft Academic Search

Gentian violet (GV) is recommended for initial treatment of oral candidiasis in HIV-infected patients in resource-limited\\u000a settings. Currently GV is not used because of its staining effects. In this study, we investigated the staining capacity of\\u000a three different concentrations of GV to determine a concentration that does not cause staining. The selected concentration\\u000a that did not cause staining was evaluated

R. J. Jurevic; R. S. Traboulsi; P. K. Mukherjee; R. A. Salata; M. A. Ghannoum

2011-01-01

438

Fibre sizes and histochemical staining characteristics in normal and chronically stimulated fast muscle of cat.  

PubMed

1. Normal and chronically stimulated peroneus longus muscles of the cat's hind limb were studied with respect to fibre size and staining properties for myofibrillar (myosin) adenosine triphosphatase (ATPase) and succinate dehydrogenase (SDH) activity. The intensity of staining for SDH activity was measured by microphotometry from the central portions of the muscle fibres ('core-SDH staining'). For comparison, histochemical properties were also studied in non-stimulated soleus muscles. 2. On account of the pH sensitivity of their myofibrillar ATPase, about 18% of the fibres in normal peroneus longus muscles were classified as type I, and about half of the remainder as II A and II B respectively. 3. In the normal peroneus longus muscles, the mean diameter of single muscle fibres generally varied between about 25 and 75 micron, whereby the average size of type I less than type II. 4. In the normal peroneus longus muscles the staining intensity for core SDH varied over a wide range. The average heaviness of staining was clearly ranked in the order type I greater than type II A greater than type II B. 5. Chronic stimulation was given to the deafferented common peroneal nerve by aid of a portable and remotely controlled mini-stimulator. The stimulation was delivered in 'tonic' patterns (greater than or equal to 50% of total time taken up by activity) of 'fast' (20 or 40 Hz) or 'slow' (5 or 10 Hz) rates. 6. Prior to the period of long-term stimulation, the cats had been subjected to a dorsal rhizotomy and hemispinalization on the ipsilateral (left) side. In the absence of chronic stimulation, these operations had no evident effects on the sizes or staining properties of peroneus longus fibres. 7. After 8 weeks of treatment with tonic patterns of stimulation, the fibres of peroneus longus muscles clearly became more similar to each other with respect to their diameter as well as their staining for ATPase and SDH activity. With respect to ATPase staining, however, the chronically stimulated peroneus longus fibres had become more similar to non-stimulated soleus fibres than to non-stimulated type I fibres of peroneus longus. With respect to the staining for core SDH, the chronically stimulated fibres all became similar to normal II A fibres of peroneus longus. The 'fast' and 'slow' patterns of chronic stimulation had the same effects on the staining properties. 8. Chronically stimulated peroneus longus muscles showed a decrease in fibre diameter which corresponded, roughly, to the concomitant decrease in muscle weight.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:2957493

Donselaar, Y; Eerbeek, O; Kernell, D; Verhey, B A

1987-01-01

439

Fibre sizes and histochemical staining characteristics in normal and chronically stimulated fast muscle of cat.  

PubMed Central

1. Normal and chronically stimulated peroneus longus muscles of the cat's hind limb were studied with respect to fibre size and staining properties for myofibrillar (myosin) adenosine triphosphatase (ATPase) and succinate dehydrogenase (SDH) activity. The intensity of staining for SDH activity was measured by microphotometry from the central portions of the muscle fibres ('core-SDH staining'). For comparison, histochemical properties were also studied in non-stimulated soleus muscles. 2. On account of the pH sensitivity of their myofibrillar ATPase, about 18% of the fibres in normal peroneus longus muscles were classified as type I, and about half of the remainder as II A and II B respectively. 3. In the normal peroneus longus muscles, the mean diameter of single muscle fibres generally varied between about 25 and 75 micron, whereby the average size of type I less than type II. 4. In the normal peroneus longus muscles the staining intensity for core SDH varied over a wide range. The average heaviness of staining was clearly ranked in the order type I greater than type II A greater than type II B. 5. Chronic stimulation was given to the deafferented common peroneal nerve by aid of a portable and remotely controlled mini-stimulator. The stimulation was delivered in 'tonic' patterns (greater than or equal to 50% of total time taken up by activity) of 'fast' (20 or 40 Hz) or 'slow' (5 or 10 Hz) rates. 6. Prior to the period of long-term stimulation, the cats had been subjected to a dorsal rhizotomy and hemispinalization on the ipsilateral (left) side. In the absence of chronic stimulation, these operations had no evident effects on the sizes or staining properties of peroneus longus fibres. 7. After 8 weeks of treatment with tonic patterns of stimulation, the fibres of peroneus longus muscles clearly became more similar to each other with respect to their diameter as well as their staining for ATPase and SDH activity. With respect to ATPase staining, however, the chronically stimulated peroneus longus fibres had become more similar to non-stimulated soleus fibres than to non-stimulated type I fibres of peroneus longus. With respect to the staining for core SDH, the chronically stimulated fibres all became similar to normal II A fibres of peroneus longus. The 'fast' and 'slow' patterns of chronic stimulation had the same effects on the staining properties. 8. Chronically stimulated peroneus longus muscles showed a decrease in fibre diameter which corresponded, roughly, to the concomitant decrease in muscle weight.(ABSTRACT TRUNCATED AT 400 WORDS) Images Fig. 1 Fig. 3 Fig. 5 PMID:2957493

Donselaar, Y; Eerbeek, O; Kernell, D; Verhey, B A

1987-01-01

440

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

PubMed

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

Deng, M Y; Cliver, D O

1992-08-01