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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

Applicability of the fluorescein diacetate method of detecting active bacteria in freshwater  

Microsoft Academic Search

Fluorescein diacetate (FDA) hydrolysis was evaluated as a means to detect actively metabolizing bacteria in freshwater. Fluorescein diacetate, a nonfluorescent derivative of fluorescein, can be transported across cell membranes and deacetylated by nonspecific esterases. Resultant fluorescein accumulates within cells and allows direct visualization by epifluorescent microscopy. Application of FDA to a variety of freshwater habitats yielded estimates of active cells

Thomas H. Chrzanowski; Rhonda D. Crotty; James G. Hubbard; Robert P. Welch

1984-01-01

3

Fluorescein diacetate hydrolysis as an estimator of microbial biomass on coniferous needle surfaces  

Microsoft Academic Search

Estimating microbial standing crops and microbial production in natural habitats has been difficult for microbial ecologists. The present paper describes a simple spectrophotometric assay based on the hydrolysis of fluorescein diacetate which estimates well the standing crops of microbial cells on coniferous needles and twigs. A technique is also presented for correlating optical density readings with actual dry weights of

Ronald Swisher; George C. Carroll

1980-01-01

4

Fluorescein diacetate hydrolysis as a measure of microbial activity in aquatic systems: Application to activated sludges  

Microsoft Academic Search

Fluorescein diacetate (FDA) hydrolysis has mainly been used, in soil studies, for measurement of microbial activity and\\/or for enumeration of bacteria. A protocol is proposed to apply the method to sewage treatment plant activated sludge. The results are compared with values of ETS (electron transport system) activity and oxygen consumption. Unlike ETS activity, FDA hydrolysis is not expected to be

D. A. Fontvieille; A. Outaguerouine; D. R. Thevenot

1992-01-01

5

Fluorescein-Labeled beta-Glucosidase as a Bacterial Stain.  

National Technical Information Service (NTIS)

Beta-glucosidase labeled with fluorescein isothiocyanate was used as a simple staining reagent with selected gram-positive and gram-negative organisms. Staining appeared to be dependent on the presence of accessible glycosidic-type linkages as well as the...

A. Pital S. L. Janowitz C. E. Hudak E. E. Lewis

1967-01-01

6

EPIDERMAL INTERCELLULAR STAINING WITH FLUORESCEIN-CONJUGATED PHYTOHEMAGGLUTININS  

Microsoft Academic Search

Fluorescein-conjugated phytohemagglutinins, Concanavalin A and Phytohemagglutinin P were used to stain frozen-cut sections of human skin to determine the presence and distribution of carbohydrate-containing substances in the epidermis. An intercellular staining pattern was observed which appeared identical to that found in various forms of pemphigus. The intercellular staining could be inhibited, or abolished, by ?-methyl-D-mannopyranoside or ?-methyI-D-glucopyranoside in the case

Michael L. Nieland

1973-01-01

7

Fluorescein-labeled ?-Glucosidase as a Bacterial Stain  

PubMed Central

Fluorescein isothiocyanate-labeled ?-glucosidase was used as a simple staining reagent with selected gram-positive and gram-negative organisms. Staining in situ appeared to be dependent on the presence of accessible glycosidic-type linkages in the bacterial cell wall. Extensive wall damage or lysis did not occur when stained cells were suspended in washing and mounting solutions. The apparent specificity of labeled enzyme for wall substance was tested by blocking reactions, staining of isolated cell walls, and failure to stain substances lacking appropriate glycosidic linkages. Severe cell wall lesions were produced after prolonged contact with labeled enzyme, and this phenomenon may also be related to staining specificity. Gram-negative organisms and spores were poorly stained unless protected glycopeptide substrate was previously exposed by treatment of cells with thioglycolic acid or dilute alkaline sodium hypochlorite solution. A potential for staining tissues and cell lines may also exist. Some possible applications of labeled enzymes are briefly discussed. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4

Pital, Abe; Janowitz, Sheldon L.; Hudak, Charles E.; Lewis, Evelyn E.

1967-01-01

8

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

SciTech Connect

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

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

1987-01-01

9

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

10

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

11

Which fluorophore is brightest? A comparison of the staining obtained using fluorescein, tetramethylrhodamine, lissamine rhodamine, texas red, and cyanine 3.18  

Microsoft Academic Search

There are several red-emitting fluorophores available for immunofluorescence studies, including tetramethylrhodamine, lissamine rhodamine, Texas Red, and cyanine 3.18; however, it is unclear which of these is best. The present study compared the brightness of these fluorophores to that of fluorescein. Staining was attempted using a primary antibody raised against serotonin and a secondary antibody that was conjugated with either fluorescein

M. W. Wessendorf; T. C. Brelje

1992-01-01

12

Pin-printing utility and feature characteristics on planar surfaces & mechanism of fluorescein corneal staining  

NASA Astrophysics Data System (ADS)

This thesis covers two areas of research both of which have spectroscopy at the center. The first part of the thesis investigates pin-printing strategies on planar surfaces. These surfaces include silicon based substrates such as glass microscope slides, mirrored glass slides, and oxidized porous silicon. Spectroscopy techniques are used to assess the results of pin-printed microarrays on these substrates. Topics investigated include stability, uniformity, and basic screening of the materials printed onto these surfaces. The second part of this thesis focuses on elucidating the mechanism behind solution induced corneal staining. Spectroscopy experiments are reported that investigate specific interactions that are occurring at the human corneal epithelial cell surface. Different interactions are probed by using molecular fluorescence spectroscopy.

Kraut, Nadine D.

13

Fluorescein. Physiochemical factors affecting its fluorescence.  

PubMed

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

Romanchuk, K G

1982-01-01

14

RED Facts: Chlorhexidine Diacetate.  

National Technical Information Service (NTIS)

The fact sheet summarizes the information in the RED document for reregistration case 3038, chlorhexidine diacetate. Chlorhexidine diacetate is a disinfectant used to control bacteria on agricultural premises, egg handling and packing equipment, and meat ...

1996-01-01

15

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

16

Histologic Localization of Sodium Fluorescein in Human Ocular Tissues.  

National Technical Information Service (NTIS)

The distribution pattern of sodium fluorescein in human eyes microscopically was studied. The ciliary body showed early and diffuse leakage, with staining of the basement membrane of the nonpigmented ciliary epithelium, indicating movement of fluorescein ...

R. T. McMahon M. O. M. Tso I. W. McLean

1975-01-01

17

Fluorescein Diacetate: A Potential Biological Indicator for Arid Soils  

Microsoft Academic Search

A field study was undertaken to identify a potential biological indicator for arid soils. The study area covered five districts (111,681km) in which annual rainfall varied from 217 to 427mm and soil texture ranged from sandy to clay loam. The surface 30cm of arid soil from agricultural field sites differing in soil properties and cropping pattern were used in the

G. K. Aseri; J. C. Tarafdar

2006-01-01

18

21 CFR 182.6197 - Calcium diacetate.  

Code of Federal Regulations, 2013 CFR

... 2013-04-01 2013-04-01 false Calcium diacetate. 182.6197 Section 182.6197...RECOGNIZED AS SAFE Sequestrants 1 § 182.6197 Calcium diacetate. (a) Product. Calcium diacetate. (b) Conditions of use....

2013-04-01

19

21 CFR 582.6197 - Calcium diacetate.  

Code of Federal Regulations, 2013 CFR

... 2013-04-01 2013-04-01 false Calcium diacetate. 582.6197 Section 582.6197...RECOGNIZED AS SAFE Sequestrants 2 § 582.6197 Calcium diacetate. (a) Product. Calcium diacetate. (b) Conditions of use....

2013-04-01

20

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

21

Reregistration Eligibility Decision (RED): Chlorohexidine Diacetate. (Includes RED Facts: Chlorhexidine Diacetate Fact Sheet).  

National Technical Information Service (NTIS)

This document presents the Agency's decision regarding the reregistration eligibility of the registered uses of chlorhexidine diacetate. Section I is the introduction. Section II describes chlorhexidine diacetate, its uses, data requirements and regulator...

1996-01-01

22

21 CFR 522.1078 - Gonadorelin diacetate tetrahydrate.  

Code of Federal Regulations, 2010 CFR

...2010-04-01 false Gonadorelin diacetate tetrahydrate. 522.1078 Section 522.1078...522.1078 Gonadorelin diacetate tetrahydrate. (a) Specifications . Each...micrograms (µg) of gonadorelin diacetate tetrahydrate. (b) Sponsors . See Nos....

2010-04-01

23

21 CFR 522.1078 - Gonadorelin diacetate tetrahydrate.  

Code of Federal Regulations, 2010 CFR

...2009-04-01 false Gonadorelin diacetate tetrahydrate. 522.1078 Section 522.1078...522.1078 Gonadorelin diacetate tetrahydrate. (a) Specifications . Each...micrograms (µg) of gonadorelin diacetate tetrahydrate. (b) Sponsors . See Nos....

2009-04-01

24

Fluorescein Derivatives in Intravital Fluorescence Imaging  

PubMed Central

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

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

2013-01-01

25

Fluorescein angiography findings in phacomatosis pigmentovascularis.  

PubMed

The authors report the fluorescein angiography findings in a 3-month-old patient with phacomatosis cesioflammea, which revealed venous-venous anastomoses in addition to previously undescribed features of peripheral retinal vascular nonperfusion. The authors encourage physicians to consider phacomatosis pigmentovascularis in the differential diagnosis of patients presenting with facial port-wine stain and to screen these patients for peripheral retinal avascularity in addition to glaucoma and primary uveal melanoma. PMID:23413944

Henry, Christopher R; Hodapp, Elizabeth; Hess, Ditte J; Blieden, Lauren S; Berrocal, Audina M

2013-01-01

26

Variation in human islet viability based on different membrane integrity stains.  

PubMed

Membrane integrity fluorescent staining is used routinely to evaluate islet viability. Results are used as one of the determining factors in islet product release criteria, and are used to assess the efficacy of different culture conditions. Recently, it has been observed that there is variation in the viability staining of freshly isolated islets based on which viability assay is used. This investigation compares three membrane integrity stains for the viability assessment of isolated human islets. Fluorescein diacetate/propidium iodide (FDA/ PI), the current standard method for assessing islet viability, demonstrates intense extracellular fluorescence, reducing the differential staining of intact islets. We further evaluated SYTO-13/ethidium bromide (SYTO/ EB) and calcein AM/ethidium homodimer (C/EthD) as alternative viability assays, and found considerable variation between FDA/PI and either SYTO/EB or C/EthD staining. Preparations of human islets were obtained from cadaveric pancreata after collagenase digestion, mechanical separation, and purification by continuous Ficoll gradient centrifugation. For each preparation, two replicate samples of 50 islets were counted for each stain, and the percent viability calculated. The results for SYTO/EB and C/EthD were nearly identical [57.6 +/- 7.3% and 57.9 +/- 7.2%, respectively (mean +/- SEM), N = 11]. FDA/PI-stained islets, however, showed consistently elevated values when compared to SYTO/EB. Accurate assessment of islet viability remains a critical determinant of islet product release. The discrepancies found between FDA/PI scoring and visual quality, compared with alternative stains, suggests that the FDA/PI stain may not be the optimal approach to assess islet viability. PMID:15565860

Barnett, M J; McGhee-Wilson, D; Shapiro, A M J; Lakey, J R T

2004-01-01

27

Effects of Chlorhexidine Diacetate on Ruminal Microorganisms  

Microsoft Academic Search

. The objectives of this study were to examine the effects of chlorhexidine diacetate on growth and L-lactate production by\\u000a Streptococcus bovis JB1 as well as the effects of this antimicrobial compound on the mixed ruminal microorganism fermentation. Addition of 1.8\\u000a ?M chlorhexidine diacetate to glucose medium resulted in a lag in growth by S. bovis JB1, and growth was

Salah A. Attia-Ismail; Scott A. Martin

1998-01-01

28

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.

29

Seidel's test using 10% fluorescein.  

PubMed

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

Romanchuk, K G

1979-10-01

30

STUDIES ON FLUORESCENT ANTIBODY STAINING  

PubMed Central

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

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

1961-01-01

31

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

Federal Register 2010, 2011, 2012, 2013

...use in diagnostic fluorescein angiography or angioscopy of the retina and iris vasculature. AK-FLUOR (fluorescein sodium injection...use in diagnostic fluorescein angiography or angioscopy of the retina and iris vasculature. FUNDUSCEIN-25 (fluorescein...

2012-04-18

32

Intracarotid Fluorescein Angiography  

PubMed Central

Patterns of blood flow were examined in the surface vessels of the surgically exposed brain by intracarotid injection of 1% fluorescein and rapid serial photographs timed by a photo-cell signal. Matching colour filters were used for black and white or Ektachrome film. As developed in cats and monkeys, and applied in five patients during craniotomy, the technique gave a picture of flow patterns in the pial and cortical vascular bed, demonstrating water-shed areas bordering major arterial territories, laminar flow in veins, and, in particular, the details of filling and clearing in the fine pial vessels, the superficial cortical capillary bed and in the vascular beds of tumours. Since these features are rendered in finer detail and sharper contrast than by standard x-ray angiography, the method affords a new means of more adequately examining the epicerebral circulation in man during craniotomy for a variety of lesions. ImagesFig. 1Fig. 2aFig. 2bFig. 3Fig. 4Fig. 5Fig. 6

Feindel, William; Yamamoto, Y. Lucas; Hodge, Charles P.

1967-01-01

33

Dihydrofluorescein diacetate is superior for detecting intracellular oxidants: comparison with 2?,7?-dichlorodihydrofluorescein diacetate, 5(and 6)-carboxy-2?,7?-dichlorodihydrofluorescein diacetate, and dihydrorhodamine 123  

Microsoft Academic Search

To detect intracellular oxidant formation during reoxygenation of anoxic endothelium, the oxidant-sensing fluorescent probes, 2?,7?-dichlorodihydrofluorescein diacetate, dihydrorhodamine 123, or 5(and 6)-carboxy-2?,7?-dichlorodihydrofluorescein diacetate were added to human umbilical vein endothelial cells during reoxygenation. None of these fluorescent probes were able to differentiate the controls from the reoxygenated cells in the confocal microscope. However, dihydrofluorescein diacetate demonstrated fluorescence of linear structures, consistent

STEPHEN L. HEMPEL; GARRY R. BUETTNER; Yunxia Q OMalley; DUANE A. WESSELS; DAWN M. FLAHERTY

1999-01-01

34

Factors That Affect Fluorescein Analysis.  

National Technical Information Service (NTIS)

Quality assurance aspects are of considerable consequence in experimental studies. Because of the widespread use of fluorescein as an analytical tracer in aerosol studies, a set of experiments was conducted that summarize the effects of parameters that in...

J. Kesavan R. W. Doherty D. G. Wise A. McFarland

2001-01-01

35

FLUORESCEIN-CONJUGATED BOVINE ALBUMIN  

PubMed Central

Fluorescein-bovine albumin conjugates have been prepared and found not to differ appreciably in size, shape, and homogeneity from the precursor, bovine serum albumin. Fluorescein has also been conjugated to rat plasma proteins. Their disappearance rates from the circulation of rats correspond with those obtained from the use of isotope labeling. Their sites of localization in rat tissues were shown to be in the cytoplasm but not in the nuclei of Kupffer cells, fixed macrophages, granulocytes, and proximal renal tubules. Adsorption to endothelium was a characteristic finding. Extracellular localizations were predominantly in the lumina of blood vessels and proximal renal tubules (but never in the lumina of collecting tubules), and the interstitial fluid of skeletal and cardiac muscle (but not that of glandular organs such as the adrenals, liver, and spleen). BAC absorption from the skin of rabbits requires days whereas sodium fluorescein absorption is measured in hours, attesting to the persistence of the colloidal state of BAC in vivo. Fluorescein conjugates have been used to visualize the transcapillary passage of circulating proteins in the mesenteric circulation of frogs and rats by direct microscopic observation and found to diffuse slowly in the manner predicted for plasma proteins. The normal cutaneous vessels of the rat are impermeable in the gross to the labeled proteins; second degree burn promptly increases the permeability of these vessels rendering the presence of the label detectable in the gross in the skin. The process of labeling does not render guinea pig albumin antigenic, although slight antigenicity results from labeling whole plasma protein. It is believed that sufficient biological evidence is presented to support the conclusion that fluorescein-conjugated plasma proteins, particularly albumin, behave in vivo like their native precursors.

Schiller, Alfred A.; Schayer, Richard W.; Hess, E. L.

1953-01-01

36

Gram Stain  

MedlinePLUS

... an infected site are the most commonly performed microbiology tests used to identify the cause of a ... 2012) Cavanaugh D, Keen M, American Society for Microbiology. The Gram Stain: An Animated Approach. Available online ...

37

Negative Staining  

NSDL National Science Digital Library

This video from CUNY Kingsborough Community College describes negative staining. The brief demonstration is described step by step and would be easy to replicate in a laboratory setting. Running time for the video is 1:32.

2013-06-21

38

Experimental carotid occlusion: funduscopic and fluorescein angiographic findings.  

PubMed Central

A characteristic fundus picture was consistently produced following acute bilateral common carotid artery ligation in mature rats, reminiscent of human carotid occlusive disease. Two days after ligation it consisted of dilatation and tortuosity of retinal veins, blurring and swelling of the optic disc, retinal whitening primarily along the venous distribution, and straightening of retinal arteries. Fluorescein angiography showed hyperfluorescence of the disc, delay in the rate of retinal arterial and venous filling, venous dilatation, disc oedema, disruption of the retinal capillary bed pattern, and late peripapillary staining/leakage. This pattern was not seen in rats which underwent acute unilateral ligation, although some mild changes were seen on fluorescein angiography. The vascular alterations seemed to regress spontaneously within one week. A peripapillary 'halo' and a granular-appearing nerve fibre layer developed later, exclusively in bilaterally ligated animals. Images

Spertus, A D; Slakter, J S; Weissman, S S; Henkind, P

1984-01-01

39

pH Fluorescent Probes: Chlorinated Fluoresceins  

Microsoft Academic Search

A series of regiospecific chlorinated fluoresceins have been synthesized by the reaction of the regiospecific chlorinated\\u000a resorcinols with chlorinated phthalic anhydride. The regioisomers were successfully separated by chromatography. The photophysical\\u000a properties of the obtained chlorinated fluoresceins were examined and found their absorption and emission maxima at long wavelength\\u000a with high fluorescence quantum yield. Especially, pH-dependent properties of chlorinated fluoresceins have

Feng-Yan Ge; Li-Gong Chen

2008-01-01

40

SLE retinopathy: evaluation by fluorescein angiography.  

PubMed Central

Fifty-two patients with systemic lupus erythematosus (SLE) were examined by fluorescein angiography, and retinopathy was detected in 15. Three patterns of retinopathy were discerned: 4 patients had disc vasculitis, 6 had multiple cotton-wool spots, and 5 had a normal fundal appearance but fluorescein leakage on angiography. One patient had arterial occlusive disease with retinal neovascularisation and another had extensive venous disease. With 3 exceptions retinopathy was found only in patients with active SLE. No association was discovered between retinopathy and cerebral disease; in particular, fluorescein angiography did no assist the diagnosis of mild cerebral lupus. Images

Lanham, J G; Barrie, T; Kohner, E M; Hughes, G R

1982-01-01

41

Microgels and ionic associations in solutions of cellulose diacetate  

Microsoft Academic Search

Solutions of cellulose diacetate (CDA) from two sources (cotton linters and wood pulp Floranier) were analysed in various solvents by size exclusion chromatography (SEC). Without special precautions, the SEC chromatograms presented three peaks or prehumps before the main polymer peak. The first prehump which could be eliminated by ultracentrifugation corresponded to microgels whose sugar composition was determined. These

E. Fleury; J. Dubois; C. Lonard; J. P. Joseleau; H. Chanzy

1994-01-01

42

Light-induced transformations of hematoporphyrin diacetate and hematoporphyrin.  

PubMed

Illumination of hematoporphyrin diacetate (HP-Diac) and hematoporphyrin (HP) in phosphate-buffered solutions causes the formation of a stable photoproduct absorbing at 636 nm. Simultaneously the photo-oxidation of HP-Diac and HP takes place. These phototransformations depend on the illumination dose and the concentration of the HP-Diac or HP solution, and cause qualitatively the same spectral changes independently of the light source used. PMID:3148699

Rotomskiene, J; Kapociute, R; Rotomskis, R; Jonusauskas, G; Szit, T; Nizhnik, A

1988-11-01

43

Joint fluid Gram stain  

MedlinePLUS

Gram stain of joint fluid ... result means no bacteria are present on the Gram stain. Note: Normal value ranges may vary slightly ... Abnormal results mean bacteria were seen on the Gram stain. This may be a sign of a ...

44

Video fluorescein angiography: Method and clinical application  

Microsoft Academic Search

Video fluorescein angiography, combined with a picture analyzing system, is a clinically applicable, objective method of evaluating the retinal blood-flow parameters. Optical density measurements were performed on videorecordings of fluorescence angiograms by means of a picture-analyzing system in order to determine the circulation parameters of the retina. These included: the arm-retina time (ART), the arteriovenous passage time (AVP), and the

S. Wolf; F. Jung; H. Kiesewetter; N. Krber; M. Reim

1989-01-01

45

Absorption and fluorescence properties of fluorescein  

Microsoft Academic Search

We have characterized the protolytic equilibria of fluorescein and determined the spectroscopic properties of its protolytic forms. The protolytic constants relating the chemical activities (which at low ionic strength equal concentrations) of the cation, neutral form, anion and dianion are pK1 = 2.08, pK2 = 4.31, and pK3 = 6.43. All forms have rather high molar absorptivities being ?437FH31 =

Robert Sjback; Jan Nygren; Mikael Kubista

1995-01-01

46

Antihistamines as prophylaxis against side reactions to intravenous fluorescein.  

PubMed Central

Systemic antihistamines were administered prior to dye injection in 50 patients undergoing fluorescein angiography. The patients were monitored for side effects. Venous blood samples were obtained before and at three, ten and thirty minutes after intravenous administration of sodium fluorescein and analyzed for histamine levels. Three patients (6%) developed minor side effects of nausea or dizziness; this compares to an incidence of 21% in a previous series of patients from our institution untreated with antihistamines. A three-fold increase in plasma histamine levels occurred in 28% of patients following fluorescein and antihistamine injection; this compares with a 26% incidence of increase in plasma histamine in patients receiving fluorescein without antihistamines (as determined in a previous study). Prophylactic antihistamines should be considered in patients undergoing fluorescein angiography if they have a history of previous allergies or side reactions during prior fluorescein studies. However, complete prophylaxis against severe side reactions to fluorescein injections is not assured with antihistamines.

Ellis, P P; Schoenberger, M; Rendi, M A

1980-01-01

47

Port-Wine Stain  

MedlinePLUS

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

48

[Amyloid staining. III].  

PubMed

Anisotrophy of collagen fibres can be abolished, without change of the anomalous colour of amyloid, by mounting Congo-red stained sections in glycerin. Treatment with sodium hydroxide solution prevents staining of secondary amyloid with Sirius red. PMID:2466382

Krutsay, M

1988-01-01

49

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 ...

50

Fluorescein-labeled wheat germ agglutinin stains the pine wood nematode, Bursaphelenchus xylophilus  

Microsoft Academic Search

Pine wilt disease, caused by the pine wood nematode (PWN, Bursaphelenchus xylophilus), is a major threat to pine forests throughout East Asia. Nonetheless, its mechanism of invasion has not yet been described\\u000a in detail. To better understand the pathology of this disease, it is important to examine the distribution of PWNs within\\u000a pine tissue during the course of disease development.

Masabumi Komatsu; Jounga Son; Norihisa Matsushita; Taizo Hogetsu

2008-01-01

51

Gram stain of urethral discharge  

MedlinePLUS

Urethral discharge Gram stain ... microscope slide. A series of stains called a gram stain is applied to the specimen. The stained ... culture ) should be performed in addition to the gram stain. More sophisticated diagnostic tests (such as PCR ...

52

Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy.  

PubMed Central

An investigation on the photobleaching behavior of fluorescein in microscopy was carried out through a systematic analysis of photobleaching mechanisms. The individual photochemical reactions of fluorescein were incorporated into a theoretical analysis and mathematical simulation to study the photochemical processes leading to photobleaching of fluorescein in microscopy. The photobleaching behavior of free and bound fluorescein has also been investigated by experimental means. Both the theoretical simulation and experimental data show that photobleaching of fluorescein in microscopy is, in general, not a single-exponential process. The simulation suggests that the non-single-exponential behavior is caused by the oxygen-independent, proximity-induced triplet-triplet or triplet-ground state dye reactions of bound fluorescein in microscopy. The single-exponential process is a special case of photobleaching behavior when the reactions between the triplet dye and molecular oxygen are dominant.

Song, L; Hennink, E J; Young, I T; Tanke, H J

1995-01-01

53

Simplified method for DNA and protein staining of human hematopoietic cell samples. [Cell flow systems  

SciTech Connect

A rapid reproducible method yielding high resolution analysis of DNA and protein in human hematopoietic cell samples has been developed by modification of the propidium iodide and fluorescein isothiocyanate procedure. Cell staining involves sequential addition of each reagent (RNase, fluorescein isothiocyanate and propidium iodide) to ethanol-fixed cells and requires no centrifugation steps. Stained cells are analyzed in the reagent solutions. Analysis of bone marrow samples from multiple myeloma patients showed mixed normal and aneuploid populations with a major portion of the aneuploid cells having a significantly higher protein content. This approach permitted differential cell cycle analysis of normal and the aneuploid populations.

Crissman, H.A. (Los Alamos National Lab., NM); Egmond, J.V.; Holdrinet, R.S.; Pennings, A.; Haanen, C.

1981-01-01

54

THE DISTRIBUTION OF MUSCLE ANTIGENS IN CONTRACTED MYOFIBRILS DETERMINED BY FLUORESCEIN-LABELED ANTIBODIES  

PubMed Central

Chick myofibrils in different states of contraction were treated with fluorescein-labeled antibodies. The rabbit antibodies were prepared against chick myosin, light and heavy meromyosins, and actin. For any one state of contraction, a single myofibril was photographed through the phase contrast microscope, stained with one of the antisera, and photographed through the fluorescence microscope. The cytological changes in the sarcomeres accompanying contraction as observed under phase were correlated with changes in the distribution of the precipitated antibodies as observed under the fluorescence microscope. The changing patterns observed through the fluorescence microscope were compared with those predicted by the sliding filament model of contraction.

Tunik, Bernard; Holtzer, Howard

1961-01-01

55

Sensitivity of detecting Chlamydia trachomatis elementary bodies in smears by use of a fluorescein labelled monoclonal antibody: comparison with conventional chlamydial isolation  

Microsoft Academic Search

Commercially produced fluorescein labelled monoclonal antibodies for the detection of Chlamydia trachomatis have recently become available. One is for detecting inclusions in cell culture (culture confirmation) and the other for detecting elementary bodies in smears from potentially infected sites. We have compared the two monoclonal antibodies with our routine isolation method, which utilises Giemsa staining of cycloheximide treated McCoy cell

B J Thomas; R T Evans; D A Hawkins; D Taylor-Robinson

1984-01-01

56

A standardized visual scale for evaluation of tear fluorescein clearance  

Microsoft Academic Search

Objective: To evaluate the correlation and the agreement between a validated fluorometric technique (fluorescein clearance test) and a newly developed, clinically practical standardized visual scale to evaluate tear fluorescein clearance. Also, the ability of this new method to distinguish healthy persons from patients reporting ocular irritation associated with meibomian gland disease (MGD), aqueous tear deficiency (ATD), or both was tested.

Angelo Macri; Maurizio Rolando; Stephen Pflugfelder

2000-01-01

57

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

58

Candida, fluorescent stain (image)  

MedlinePLUS

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

59

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.

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

2013-01-01

60

Westphalen's diol diacetate: 19(10->5)-abeo-5?-cholest-9-ene-3?,6?-diyl diacetate  

PubMed Central

The structure of the title steroid [alternative name: 3?,6?-diacetoxy-5?-methyl-19-norcholest-9(10)-ene], C31H50O4, confirms the generally accepted mechanism for the rearrangement of a cholestan-5?-ol derivative reported a century ago by Westphalen. The methyl group at position 10 of the starting material migrates to position 5 in the steroidal nucleus, while a ?9 bond is formed, as indicated by the C=C bond length of 1.347?(4)?. The methyl transposition leaves the 5R configuration unchanged, with the methyl oriented towards the ? face. During the rearrangement, the steroidal B ring experiences a conformational distortion from chair to envelope with the C atom at position 6 as the flap. In the title structure, the isopropyl group of the side chain is disordered over two positions, with occupancies of 0.733?(10) and 0.267?(10). The carbonyl O atom in the acetyl group at C3 is also disordered with an occupancy ratio of 0.62?(4):0.38?(4).

Ramirez Hernandez, Johana; Sandoval-Ramirez, Jesus; Meza-Reyes, Socorro; Vega Baez, Jose Luis; Bernes, Sylvain

2012-01-01

61

Apparatus Would Stain Microscope Slides  

NASA Technical Reports Server (NTRS)

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

Breeding, James D.

1993-01-01

62

Use of fluorescein angiography in incontinentia pigmenti: a case report.  

PubMed

The authors present the case of a 6-month-old female infant with a known diagnosis of incontinentia pigmenti with a normal examination by indirect ophthalmoscopy. However, fluorescein angiography revealed vascular abnormalities that were not detected by indirect ophthalmoscopy. Follow-up examination revealed progressive vascular changes that again were only detectable by fluorescein angiography. Because vision loss can cause significant morbidity in incontinentia pigmenti, the use of fluorescein angiography as an adjunctive tool with exams under anesthesia may provide invaluable information in the detection of early vascular changes in this disease. PMID:23410815

Tzu, Jonathan H; Murdock, Jennifer; Parke, D Wilkin; Warman, Roberto; Hess, Ditte J; Berrocal, Audina M

2013-01-01

63

Antibacterial polyurethane nanocomposites using chlorhexidine diacetate as an organic modifier.  

PubMed

Polymer nanocomposites (NCs) are hypothesised to have enhanced barrier properties compared with pristine polymer, allowing more sustained drug release from the materials. In these NC systems active agents are typically incorporated into the polymer matrix and the release kinetics are theoretically perturbed by well dispersed nanoparticle inclusions. An alternative approach is to exploit active agent interactions with the nanoinclusion. In the proposed NC system, the driving hypothesis is that active agents can have dual functionality, acting as both drug and dispersant. Polyurethane-montmorillonite (PEU-MMT) NCs were prepared in which the antimicrobial agent chlorhexidine diacetate (CHX) was evaluated as an organic modifier for silicate dispersion. CHX was incorporated at various concentrations through organic modification of MMT or within the bulk polymer. X-ray diffraction and transmission electron microscopy analysis suggested that intercalated and partially exfoliated NCs were achieved, with better dispersion occurring in the presence of free CHX within the bulk. Tensile testing results showed that variations in the level of organic modification and nanoparticle loading modulated the mechanical properties. Material stiffness increased with nanoparticle loading relative to pristine PEU, and the ultimate properties decreased with nanoparticle and free CHX incorporation. Antibacterial activity against Staphylococcus epidermidis was significant in materials with higher exchanged MMT and NCs containing free CHX, for which 2-log reductions in adherent bacteria were found after 24h. CHX was successfully used to modulate the material properties in its dual role as a dispersant and antimicrobial agent, suggesting that alternative biocides of similar structure may behave comparably within PEU-MMT NC systems. PMID:20074676

Fong, N; Simmons, A; Poole-Warren, L A

2010-07-01

64

Cryo-negative staining.  

PubMed

A procedure is presented for the preparation of thin layers of vitrified biological suspensions in the presence of ammonium molybdate, which we term cryo-negative staining. The direct blotting of sample plus stain solution on holey carbon supports produces thin aqueous films across the holes, which are routinely thinner than the aqueous film produced by conventional negative staining on a continuous carbon layer. Because of this, a higher than usual concentration of negative stain (ca. 16% rather than 2%) is required for cryo-negative staining in order to produce an optimal image contrast. The maintenance of the hydrated state, the absence of adsorption to a carbon film and associated sample flattening, together with reduced stain granularity, generates high contrast cryo-images of superior quality to conventional air-dry negative staining. Image features characteristic of unstained vitrified cryo-electron microscopic specimens are present, but with reverse contrast. Examples of cryo-negative staining of several particulate biological samples are shown, including bacteriophage T2, tobacco mosaic virus (TMV), bovine liver catalase crystals, tomato bushy stunt virus (TBSV), turnip yellow mosaic virus (TYMV), keyhole limpet hemocyanin (KLH) types 1 and 2, the 20S proteasome from moss and the E. coli chaperone GroEL. Densitometric quantitation of the mass-density of cryo-negatively stained bacteriophage T2 specimens before and after freeze-drying within the TEM indicates a water content of 30% in the vitreous specimen. Determination of the image resolution from cryo-negatively stained TMV rods and catalase crystals shows the presence of optical diffraction data to ca. 10 A and 11.5 A, respectively. For cryo-negatively stained vitrified catalase crystals, electron diffraction shows that atomic resolution is preserved (to better than 20 diffraction orders and less than 3 A). The electron diffraction resolution is reduced to ca. 10 A when catalase crystal specimens are prepared without freezing or when they are freeze-dried in the electron microscope. Thin vitrified films of TMV, TBSV and TYMV in the presence of 16% ammonium molybdate show a clear indication of two-dimensional (2-D) order, confirmed by single particle orientational analysis of TBSV and 2-D crystallographic analysis of TYMV. These observations are in accord with earlier claims that ammonium molybdate induces 2-D array and crystal formation from viruses and macromolecules during drying onto mica. Three-dimensional analysis of the TBSV sample using the tools of icosahedral reconstruction revealed that a significant fraction of the particles were distorted. A reconstruction from a subset of undistorted particles produced the characteristic T = 3 dimer clustered structure of TBSV, although the spikes are shortened relative to the structure defined by X-ray crystallography. The 20S proteasome, GroEL, catalase, bacteriophage T2, TMV, TBSV and TYMV all show no indication of sample instability during cryo-negative staining. However, detectable dissociation of the KLH2 oligomers in the presence of the high concentration of ammonium molybdate conforms with existing knowledge on the molybdate-induced dissociation of this molecule. This indicates that the possibility of sample-stain interaction in solution, prior to vitrification, must always be carefully assessed. PMID:9684350

Adrian, M; Dubochet, J; Fuller, S D; Harris, J R

1998-01-01

65

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

66

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.

67

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

68

Acid-fast stain  

MedlinePLUS

... The slide is then washed with an acid solution and a different stain is applied. Bacteria that hold onto the first dye are considered "acid-fast" because they resist the acid wash. This type of bacteria is associated with tuberculosis and other infections.

69

Digitization of stained glass  

Microsoft Academic Search

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

Lindsay W. MacDonald

1997-01-01

70

Stained-Glass Pastels  

ERIC Educational Resources Information Center

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

Laird, Shirley

2009-01-01

71

Stained Glass Glue  

NSDL National Science Digital Library

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

Society, American C.

2001-01-01

72

Metal-Sulfate Induced Generation of ROS in Human Brain Cells: Detection Using an Isomeric Mixture of 5- and 6-Carboxy-2',7'-Dichlorofluorescein Diacetate (Carboxy-DCFDA) as a Cell Permeant Tracer.  

PubMed

Evolution of reactive oxygen species (ROS), generated during the patho-physiological stress of nervous tissue, has been implicated in the etiology of several progressive human neurological disorders including Alzheimer's disease (AD) and amylotrophic lateral sclerosis (ALS). In this brief communication we used mixed isomers of 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate (carboxy-DCFDA; C(25)H(14)C(l2)O(9); MW 529.3), a novel fluorescent indicator, to assess ROS generation within human neuronal-glial (HNG) cells in primary co-culture. We introduced pathological stress using the sulfates of 12 environmentally-, industrially- and agriculturally-relevant divalent and trivalent metals including Al, Cd, Cu, Fe, Hg, Ga, Mg, Mn, Ni, Pb, Sn and Zn. In this experimental test system, of all the metal sulfates analyzed, aluminum sulfate showed by far the greatest ability to induce intracellular ROS. These studies indicate the utility of using isomeric mixtures of carboxy-H(2)DCFDA diacetates as novel and highly sensitive, long-lasting, cell-permeant, fluorescein-based tracers for quantifying ROS generation in intact, metabolizing human brain cells, and in analyzing the potential epigenetic contribution of different metal sulfates to ROS-generation and ROS-mediated neurological dysfunction. PMID:22949820

Pogue, Aileen I; Jones, Brandon M; Bhattacharjee, Surjyadipta; Percy, Maire E; Zhao, Yuhai; Lukiw, Walter J

2012-01-01

73

Nickel oxide solgel films from nickel diacetate for electrochromic applications  

Microsoft Academic Search

Nickel diacetate tetrahydrate, [Ni(acetate)24H2O] and nickel diacetate dimethylaminoethanol, [Ni(acetate)2(dmaeH)2] were successfully used to deposit NiOx thin films on conductive glass substrates by solgel techniques for large area electrochromic applications. Homogeneous one layer films 100 nm thick were deposited by spin coating 0.5 M [Ni(acetate)24H2O] in dmaeH at 1000 rpm and by dip coating methods. The NiOx films were characterised by

J. L Garcia-Miquel; Q Zhang; S. J Allen; A Rougier; A Blyr; H. O Davies; A. C Jones; T. J Leedham; P. A Williams; S. A Impey

2003-01-01

74

Prelytic stimulation of target and effector cells following conjugation as measured by intracellular fluorescein fluorescence polarization  

NASA Astrophysics Data System (ADS)

The aim of the present study was to detect prelytic intracellular changes induced in target and effector cells following their conjugation at room temperature. Changes in the cytoplasmic matrix were measured by means of intracellular fluorescein fluorescence polarization (IFFP) using the Cellscan apparatus. Both natural killer and lymphocyte activated killer cells were used as effector cells, while K562 and Daudi cells lines were used as targets. The results show that following their conjugation, both the effector and the target cells show significant reductions (> 10%) in IFFP values. Changes in IFFP were induced by specific interaction and only between viable cells. No evidence of fluorescein transfer from a stained cell to its nonstained counterpart was found. To the best of our knowledge, this is the first time that effector-target interaction is monitored on an individual cell basis within a population, by means of IFFP measurements. In addition, in order to explain the physical phenomena, measurements of physical parameters which might affect the IFFP, such as changes in osmolality and pH, were performed and discussed.

Fixler, Dror; Tirosh, Reuven; Eisenthal, Avi; Lalchuk, Shlomo; Marder, Oleg; Irlin, Yosef; Deutsch, Mordechai

1998-07-01

75

In situ tracking the intracellular delivery of antisense oligonucleotides by fluorescein doped silica nanoparticles.  

PubMed

Antisense oligonucleotides (ASOs) are often utilized to interfere with gene expression at mRNA level for cancer treatment. Here, we synthesized fluorescein doped silica nanoparticles (FSNPs) and coated them by polyethyleneimine (PEI) for carrying ASOs. Agarose gel electrophoresis proved that PEI/FSNPs could load ASOs by a weight ratio as high as 30:1. We tracked the delivery process of ASOs from the ASOs/PEI/FSNPs composites to HeLa cells in situ by the confocal laser scanning microscopy (CLSM) techniques, including nuclear staining and Z-axis scanning. We found the ASOs/PEI/FSNPs composites exhibited their biological effects at specific intracellular localization, and the fluorescence of the FSNPs showed the dynamic delivery process in the cells. PMID:24913855

Zhang, Peng; Wang, Tian-Yi; Xiong, Huan-Ming; Kong, Ji-Lie

2014-09-01

76

Detection of disseminated peritoneal tumors by fluorescein diacrylate in mice  

NASA Astrophysics Data System (ADS)

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

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

2009-02-01

77

Serum and Placenta Levels of Ethynylestradiol in Presence of Ethynodiol Diacetate after Oral Administration  

Microsoft Academic Search

The ethynylestradiol concentration in the presence of ethynodiol diacetate in serum after oral administration was measured by a rapid radioimmunoassay method developed by the authors. It was found that the peak level was reached 1 h after administration, and even after 12 h a significant amount of free ethynylestradiol was present in the serum. The transfer of ethynylestradiol

J. Morvay; I. Altorjay; M. Sas

1982-01-01

78

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

79

Multiwalled carbon nanotubes inhibit fluorescein extrusion and reduce plasma membrane potential in in vitro human glioma cells.  

PubMed

In the study on the interactions of carbon nanotubes with living cells, the cell membrane deserves particular attention as it provides the first interface to initiate CNTs-cell interactions. In the present study, the inhibiting effect of multiwalled carbon nanotubes on the extrusion of fluorescein in human glioma cells was demonstrated using two procedures. To provide clues to explanation of this effect, intracellular glutathione content and reactive oxygen species production were determined as fluorescein is a specific substrate of cell membrane multidrug resistance-related protein whose transport activity requires glutathione which can be depleted under oxidative stress. The plasma membrane potential was also probed as the susceptibility of fluorescein efflux to modulation of the plasma membrane potential has been documented. Results showed a remarkable decrease in cellular glutathione level as well as an increase in reactive oxygen species production. Probe staining also indicated decreased plasma membrane potential. The data suggested that multiwalled carbon nanotubes may affect the transport activity of cell membrane multidrug resistance-related protein through reduction of intracellular glutathione content. Hypopolarization of the plasma membrane may also contribute to MWCNTs' effect. Implications of these findings are discussed. PMID:21179943

Xu, Yonghong; Chen, Xiao; Cheng, Yuli; Xing, Yiqiao

2010-06-01

80

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

81

Spectroscopy of Fluorescein (FITC) dyed colloidal silica spheres  

Microsoft Academic Search

We have measured the absorption spectrum, the emission spectrum, the emission lifetime, and the photostability of fluorescein isothiocyanate (FITC) incorporated inside colloidal silica spheres as a function of the dye concentration in the spheres, while minimizing scattering effects. Six batches of stable, monodisperse particles were synthesized with FITC up to high densities of 0.03 M. At dye concentrations above 0.001

A. Imhof; M. Megens; J. J. Engelberts; Lang de D. T. N; R. Sprik; W. L. Vos

1999-01-01

82

Serial retinal fluorescein angiography and immune therapy in Susac's Syndrome  

Microsoft Academic Search

We report a patient with Susac's disease presenting classically in a young female with an encephalopathy and visual disturbance with later deafness and tinnitus. Her encephalopathy settled, but subsequent serial fluorescein angiograms allowed sensitive monitoring of continuing sub-clinical disease activity, and provide evidence of a clear therapeutic response to immune suppression with tacrolimus (but not steroids alone) and of

Beth Mallam; Erika M. Damato; Neil J. Scolding; Clare Bailey

2009-01-01

83

Automated detection and quantification of microaneurysms in fluorescein angiograms  

Microsoft Academic Search

Fluorescein angiograms from diabetics were digitised for analysis using digital image-processing techniques. Computer algorithms were written to detect and count microaneurysms present in the images. The accuracy, speed and reproducibility of the technique were assessed and compared with those of manual counts made by clinicians from both digitised and analogue images. Free-response ROC (receiver operating characteristic) curves were used to

Timothy Spencer; Russell P. Phillips; Peter F. Sharp; John V. Forrester

1992-01-01

84

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

85

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

PubMed

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

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

2004-08-01

86

The preparation and photophysical behaviors of temperature\\/pH-sensitive polymer materials bearing fluorescein  

Microsoft Academic Search

A novel method for attaching fluorescein (via its epoxy derivate) to water soluble polymer and its temperature\\/pH-sensitive qualities of fluorescence were investigated. 3-Epoxypropoxy fluorescein (EPF) was firstly synthesized through the reaction between fluorescein and epichlorohydrin and poly (vinyl alcohol) bearing fluorescein was prepared via ring-opening reaction with EPF. Both of them were characterized by the methods of HNMR, MS, IR,

Xiaolin Guan; Xinyu Liu; Zhixing Su; Peng Liu

2006-01-01

87

Simplified method for DNA and protein staining of human hematopoietic cell samples  

SciTech Connect

A rapid reproducible method yielding high resolution analysis of DNA and protein in human hematopoietic cell samples was developed by modification of the propidium iodide (PI) and fluorescein isothiocyanate (FITC) procedure. Cell staining involved sequential addition of each reagent (RNase, FITC, and PI) to ethanol-fixed cells and requires no centrifiguation steps. Stained cells are analyzed in the reagent solutions. Analysis of bone marrow samples from multiple myeloma patients revealed mixed 2C DNA and aneuploid populations with the aneuploid cells having a significantly higher protein content. This approach permitted differential cell cycle kinetic analysis of the 2C DNA and the aneuploid population.

Crissman, H.A.; Egmond, J.V.; Holdrinet, R.S.; Pennings, A.; Haanen, C.

1980-01-01

88

Fullerenefluoresceinanthracene hybrids: a model for artificial photosynthesis and solar energy conversion  

Microsoft Academic Search

Two novel C60fluoresceinanthracene hybrids have been synthesized. Fluorescence quenching in the hybrids indicates an energy transfer from the excited state of anthracene to fluorescein and photoinduced intramolecular electron transfer from the excited state of fluorescein to the C60 moiety.

Bingwen Jing; Daoben Zhu

2004-01-01

89

Automated detection and quantification of microaneurysms in fluorescein angiograms.  

PubMed

Fluorescein angiograms from diabetics were digitised for analysis using digital image-processing techniques. Computer algorithms were written to detect and count microaneurysms present in the images. The accuracy, speed and reproducibility of the technique were assessed and compared with those of manual counts made by clinicians from both digitised and analogue images. Free-response ROC (receiver operating characteristic) curves were used to assess the performance of both the clinicians and the computer by comparing the results with "gold standards" compiled from prints of the original fluorescein angiograms. The computer performed as well as the clinicians when the latter were analysing the digitised images (512 x 512 pixel resolution), but only when one image was acquired at 4 times this resolution did the computer's performance match that of the clinicians analysing the analogue image. The automated technique was more reproducible than the manual method. PMID:1547965

Spencer, T; Phillips, R P; Sharp, P F; Forrester, J V

1992-01-01

90

Fluorescein and tetramethyl rhodamine as haptens in enzyme immunohistochemistry  

Microsoft Academic Search

Fluorescein (Fl) and tetramethyl rhodamine (Rh) were evaluated as possible candidates for a double hapten sandwich system in enzyme immunohistology. Monoclonal antibodies were raised against Fl and Rh. Their fine-specificity was tested with a competition-like assay. A pair of Mab's was selected for immunohistology in which they functioned as a bridge between Fl\\/Rh conjugated antibodies and Fl\\/Rh labeled peroxidase and

J. J. Haaijman; J. Coolen; C. J. M. Krse; G. J. Pronk; Z. F. Ming

1986-01-01

91

Diffusion and aggregation of sodium fluorescein in aqueous solutions.  

PubMed

The diffusion and aggregation of sodium fluorescein in aqueous solutions was investigated adopting density functional theory (DFT) and molecular dynamics (MD) simulations. First, DFT calculations in implicit water were used to determine minimum energy structure and atomic charges of the solute, which were then used as input for explicit water MD simulations. The self-diffusion coefficient of sodium fluorescein was calculated using the Einstein equation, computing the mean square displacement from 24 ns trajectories. The calculated diffusion coefficient, 0.42 10(-5) cm(2) s(-1), is in good agreement with literature experimental data. The simulations confirmed the tendency of fluorescein to form dimers. In order to achieve a deeper understanding of aggregation phenomena, the dimer geometry was investigated through DFT calculations both in vacuo and in implicit water using different functionals and solvation theories. The results showed that dimerization does not occur in vacuo, as charge repulsion dominates, and that the minimum energy dimer structure is symmetric and stabilized by edge-to-face ?-? interactions. The interaction energy was computed both at the DFT level and through MD simulations using Umbrella Sampling. The free interaction energy calculated with the WHAM and Umbrella Integration protocol, -1.3 kcal/mol, is in good agreement with experimental data, while the value determined using DFT calculations is significantly smaller and depends largely from the chosen functional and the computational methodology used to determine the solute-solvent boundary surface. PMID:21957875

Casalini, Tommaso; Salvalaglio, Matteo; Perale, Giuseppe; Masi, Maurizio; Cavallotti, Carlo

2011-11-10

92

Stain Resistance of Maxillofacial Materials  

Microsoft Academic Search

The resistance of three silicone and one polyvinyl chloride maxillofacial materials to staining by tea, lipstick, and disclosing solution was measured by reflectance spectrophotometry. Changes in color caused by staining were larger than changes caused by color instability of the base elastomers or pigments under conditions of accelerated aging.

A. Koran; J. M. Powers; P. J. Lepeak; R. G. Craig

1979-01-01

93

PHOSPHORUS PENTOXIDE-MONTMORILLONITE K-10 AS CATALYST FOR THE PREPARATION OF 1,1-DIACETATES UNDER SOLVENT-FREE CONDITIONS  

Microsoft Academic Search

A facile and efficient method for the preparation of 1,1-diacetates of aldehydes is improved. P2O5\\/montmorillonite K10 catalyzed 1,1-diacetates formation from aldehydes in dry media. Both aromatic and aliphatic aldehydes gave high yields (7095%) of the corresponding 1,1-diacetates. Advantages of this method are the use of an inexpensive and selective catalyst, with high yields in simple operation and short reaction time

Hossein Eshghi; Zinat Gordi

2004-01-01

94

Microscopic oxygen imaging based on fluorescein bleaching efficiency measurements.  

PubMed

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

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

2014-05-01

95

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

96

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.

2012-01-01

97

Heterogeneous catalyst for the production of ethylidene diacetate from acetic anhydride  

DOEpatents

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

Ramprasad, D.; Waller, F.J.

1998-06-16

98

Heterogeneous catalyst for the production of ethylidene diacetate from acetic anhydride  

DOEpatents

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

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

1998-01-01

99

Saliva level of ethynylestradiol in presence of ethynodiol diacetate after oral administration.  

PubMed

In the presence of ethynodiol diacetate the ethynylestradiol concentration in serum and saliva after oral administration was measured by a rapid radioimmunoassay method developed by the authors. By means of the saliva/serum quotient, and from the concentration of ethynylestradiol in the saliva samples, it was possible to infer the ethynylestradiol content of the serum. Attention is called to the relation between the quotient and time, which should be taken into account when values are calculated. 2, 4 and 12 h after oral administration, the saliva/serum quotients for an ethynylestradiol dose of 0.05 mg were 0.27, 0.76 and 0.40, respectively. PMID:6884982

Morvay, J; Altorjay, I; Sas, M

1983-01-01

100

Serial retinal fluorescein angiography and immune therapy in Susac's syndrome.  

PubMed

We report a patient with Susac's disease presenting classically in a young female with an encephalopathy and visual disturbance with later deafness and tinnitus. Her encephalopathy settled, but subsequent serial fluorescein angiograms allowed sensitive monitoring of continuing sub-clinical disease activity, and provide evidence of a clear therapeutic response to immune suppression with tacrolimus (but not steroids alone)--and of a lack of efficacy of nimodipine and aspirin. We believe this single case study has both pathogenetic and useful practical implications: the apparently favourable response to immunosuppression lends support to the hypothesis that Susac's Syndrome is an immune-mediated disease; while the presence during symptomatic clinical remission of sporadic, multi-focal episodes of hyper-fluorescence, suggestive of breakthrough vasculopathy despite treatment, underlines the fact that the natural history of this rare condition is still not fully understood. Fluorescein angiography is proposed as a sensitive and important approach to the monitoring of sub-clinical disease activity, and so optimising immune suppressive treatment. PMID:19577774

Mallam, Beth; Damato, Erika M; Scolding, Neil J; Bailey, Clare

2009-10-15

101

Movement of Fluorescein and Its Glucuronide Across Retinal Pigment Epithelium-Choroid  

Microsoft Academic Search

Purpose. To characterize movement of fluorescein and its glucuronide across the blood-retinal barrier. Methods. Retinal pigment epithelium (RPE)-choroid preparations from New Zealand albino rabbit were sealed in an Ussing-type chamber in a stabilized condition for 3 hr, where move- ment of fluorescein and fluorescein glucuronide across the RPE-choroid was studied under a short circuit condition. Results. The outward (vitreous-choroid) permeability

Satoshi Koyano; Makoto Araie; Shuichiro Eguchi

102

Intravenous fluorescein as a cause of immunoglobulin E-mediated anaphylactic shock.  

PubMed

We report a patient with severe anaphylactic shock immediately after injection of i.v. fluorescein. The patient recovered without sequela. Immunoglobulin E (IgE) mechanism was highly suggestive with significant increase in serum tryptase, positive basophil allergen threshold sensitivity (CD-sens) and histamine release tests towards fluorescein. This is, to our knowledge, the first report where CD-sens has been used to aid in diagnosing an IgE-mediated anaphylactic shock caused by fluorescein. PMID:22762373

Breidablik, A; De Pater, G H; Walther, C; Nopp, A; Guttormsen, A B

2012-09-01

103

Characterization of conformational changes in (Na,K) ATPase labeled with fluorescein at the active site  

Microsoft Academic Search

Conformational changes have been studied in (Na,K) ATPase labeled at or near the ATP binding region with fluorescein following incubation with fluorescein isothiocyanate (FITC). One or two fluorescein groups are bound per ATPase molecule. (Na,K) ATPase activity, phosphorylation from ATP, and nucleotide binding are abolished in labeled enzyme, but phosphorylation from inorganic phosphate or K-phosphatase activity are only partially inactivated.

S. J. D. Karlish

1980-01-01

104

3,6Fluorescein Diphosphate: A Sensitive Fluorogenic and Chromogenic Substrate for Protein Tyrosine Phosphatases  

Microsoft Academic Search

A highly sensitive and continuous protein tyrosine phosphatase (PTPase) assay using 3,6-fluorescein diphosphate (FDP) is described. Leukocyte phosphatase CD45 (leukocyte common antigen), protein tyrosine phosphatase-lB, and leukocyte common antigen-related protein LAR preferentially hydrolyze FDP to fluorescein monophosphate (FMP) with Vmax and Km values comparable with those of phosphotyrosine peptide substrates. Further hydrolysis of FMP to fluorescein was less efficient because

Zheng Huang; Qingping Wang; Hoa D. Ly; Arvind Gorvindarajan; John Scheigetz; Robert Zamboni; Sylvie Desmarais; Chidambaram Ramachandran

1999-01-01

105

A photostable, pH-invariant fluorescein derivative for single-molecule microscopy.  

PubMed

We herein report the comprehensive characterization of the spectral and single-photon fluorescence properties of a recently synthesized fluorescein derivative and its biotinylated analog. The fluorophore displays significant increases in photostability compared to the known fluorescein label fluorescein isothiocyanate (FITC), as well as superb pH independence. This fluorescein variant has two readily accessible functional groups (aniline NH2 and phenol OH) that can be activated or blocked independently and can serve, for instance, as a fluorescent bridge between two different recognition motifs. Excellent single-photon counting fluorescence data demonstrates that it is also a particularly appropriate probe for single-molecule studies of biological interactions. PMID:19459035

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

2009-09-01

106

Preparation, characterization, and controlled release from coprecipitates of fluorescein and magnesium hydroxide.  

PubMed

Magnesium hydroxide was precipitated as a lyophobic sol in the presence of various concentrations of fluorescein sodium (3'6'-dihydroxyspiro[isobenzofuran-1(3H),9'-[9H]xanthen++ +]-3-one, disodium salt) ranging in molar equivalents between 0.1 and 2 times that of the hydroxide. Coprecipitates were washed and dried, and release of the dye and magnesium was determined (pH 7.4, 37 degrees C) from rotating disks. Release rates varied depending upon fluorescein content. The rate of dye release was retarded by less than or equal to 10(4) times that of fluorescein sodium alone, implying the existence of some form of solid association between the components of the coprecipitates. The presence of the dye in certain concentrations reduced magnesium hydroxide dissolution rates by a factor of three. Fluorescein dissolution rates, when expressed as percent release, passed through a minimum (coincident with the dye-induced reduction in Mg(OH)2 dissolution). Adsorption experiments revealed evidence for multiaffinity binding of fluorescein at the surface of freshly precipitated Mg(OH)2. Magnesium, fluorescein, and water contents of the coprecipitates were characterized by atomic absorption and UV spectroscopy and by thermogravimetric analysis. Fluorescein content increased in direct proportion to its initial concentration in solution. Controlled, but variable release of this easily assayed dye is possible by employing precipitates with different fluorescein contents. PMID:3772746

Hickey, A J; Byron, P R

1986-08-01

107

Fluorescein angiography and fluorophotometry of the iris in pseudoexfoliation of the lens capsule  

Microsoft Academic Search

Fluorescein iris angiography and fluorophotometry were performed on a series of 9 patients with bilateral and 11 with unilateral pseudoexfoliation, 12 bilateral aphakes with pseudoexfoliation, and 7 unilateral aphakes with bilateral pseudoexfoliation. Angiography showed a loss of radial iris vessels, a heavy leak of fluorescein from the pupil margin, progressive neovascularisation of the outer 2\\/3 of the iris, and less

A. M. Brooks; W. E. Gillies

1983-01-01

108

Excretion of Fluorescein in the Urine of Women With Interstitial Cystitis  

Microsoft Academic Search

PurposeAltered bladder permeability may have a role in the pathogenesis of interstitial cystitis. Fluorescein, a fluorescent dye of molecular weight 325, has been used to assess membrane permeability. Orally ingested fluorescein normally is rapidly conjugated to glucuronate by the liver and excreted in the urine.

C. A. Tony Buffington; Bruce E. Woodworth

1997-01-01

109

Comparative Evaluation of a New Fluorescent Carboxyfluorescein Diacetate-Modified Microdilution Method for Antifungal Susceptibility Testing of Candida albicans Isolates  

Microsoft Academic Search

This report presents a fluorescent carboxyfluorescein diacetate (CFDA)-modified microdilution method used for the susceptibility testing of Candida albicans to amphotericin B, fluconazole, ketoconazole, itraconazole, voriconazole, and flucytosine. Four different broth microdilution susceptibility testing methods were simulta- neously evaluated at 24 and 48 h. The MICs determined using the CFDA-modified method (MICcfda) were compared to those obtained by the standard broth

Robert S. Liao; Robert P. Rennie; James A. Talbot

2002-01-01

110

Polarographic study of composition and stability constants of Pb(II) N-(2-acetamido) imino diacetate complexes  

Microsoft Academic Search

The complexation of Pb(II) by N-(2-acetamido) imino diacetate (ADA) has been studied polarographically (dc and ac polarographic techniques). Ac polarographic studies have been particularly helpful in deciding the reversibility of the reduction of both simple and complexed metal ions and for confirmation of the overall stability constants. A weighted least squares numerical technique has been applied for the calculation of

H. A. Azab

1992-01-01

111

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

DOEpatents

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

Ramprasad, D.; Waller, F.J.

1998-04-28

112

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

DOEpatents

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

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

1998-01-01

113

Controlled release of chlorhexidine diacetate from a porous methacrylate system: supercritical fluid assisted foaming and impregnation.  

PubMed

The release of chlorhexidine diacetate (CX) from a self-curing polymeric system based on poly(ethylmethacrylate) and tetrahydrofurfurylmethacrylate (PEM/THFM) was developed in this study. Supercritical fluid assisted impregnation and foaming was employed for preparing porous CX-PEM/THFM drug release system. X-ray diffraction (XRD) and differential scanning calorimetry (DSC) show that the crystallinity of CX significantly decreased after supercritical processing, whilst Raman spectroscopy suggested a hydrogen bonding interaction between the CX and PEM in the product. A UV-Vis dissolution study revealed that the drug release rate is almost as seven times faster in the SCF processed drug delivery system than conventional cured samples. PMID:17301965

Gong, K; Braden, M; Patel, M P; Rehman, I U; Zhang, Z; Darr, J A

2007-08-01

114

Serum and placenta levels of ethynylestradiol in presence of ethynodiol diacetate after oral administration.  

PubMed

The ethynylestradiol concentration--in the presence of ethynodiol diacetate--in serum after oral administration was measured by a rapid radioimmunoassay method developed by the authors. It was found that the peak level was reached 1 h after administration, and even after 12 h a significant amount of free ethynylestradiol was present in the serum. The transfer of ethynylestradiol into the placenta was also studied in subjects who were 10-12 weeks pregnant. Placenta/serum quotients were calculated for the ethynylestradiol, and were found to increase in parallel with the dose of the drug administered, proving that an ethynylestradiol enrichment of the placenta occurred as early as 10-12 weeks of pregnancy. PMID:6890038

Morvay, J; Altorjay, I; Sas, M

1982-01-01

115

Potentiation of Femtosecond Laser Intratissue Refractive Index Shaping (IRIS) in the Living Cornea with Sodium Fluorescein  

PubMed Central

Purpose. To assess the effectiveness of intratissue refractive index shaping (IRIS) in living corneas and test the hypothesis that it can be enhanced by increasing the two-photon absorption (TPA) of the tissue. Methods. Three corneas were removed from adult cats and cut into six pieces, which were placed in preservative (Optisol-GS; Bausch & Lomb, Inc., Irvine, CA) containing 0%, 0.25%, 1%, 1.5%, or 2.5% sodium fluorescein (Na-Fl). An 800-nm Ti:Sapphire femtosecond laser with a 100-fs pulse duration and 80-MHz repetition rate was used to perform IRIS in each piece, creating several refractive index (RI) modification lines at different speeds (between 0.1 and 5 mm/s). The lines were 1 ?m wide, 10 ?m apart, and ?150 ?m below the tissue surface. The RI change of each grating was measured using calibrated, differential interference contrast microscopy. TUNEL staining was performed to assess whether IRIS or Na-Fl doping causes cell death. Results. Scanning at 0.1 mm/s changed the RI of undoped, living corneas by 0.005. In doped corneas, RI changes between 0.01 and 0.02 were reliably achieved with higher scanning speeds. The magnitude of RI changes attained was directly proportional to Na-Fl doping concentration and inversely proportional to the scanning speed used to create the gratings. Conclusions. IRIS can be efficiently performed in living corneal tissue. Increasing the TPA of the tissue with Na-Fl increased both the scanning speeds and the magnitude of RI changes in a dose-dependent manner. Ongoing studies are exploring the use of IRIS to alter the optical properties of corneal tissue in situ, over an extended period.

Nagy, Lana J.; Ding, Li; Xu, Lisen; Knox, Wayne H.

2010-01-01

116

Extraction of Capillary Non-perfusion from Fundus Fluorescein Angiogram  

NASA Astrophysics Data System (ADS)

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

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

117

Automated segmentation of foveal avascular zone in fundus fluorescein angiography.  

PubMed

PURPOSE. To describe and evaluate the performance of a computerized automated segmentation technique for use in quantification of the foveal avascular zone (FAZ). METHODS. A computerized technique for automated segmentation of the FAZ using images from fundus fluorescein angiography (FFA) was applied to 26 transit-phase images obtained from patients with various grades of diabetic retinopathy. The area containing the FAZ zone was first extracted from the original image and smoothed by a Gaussian kernel (sigma = 1.5). An initializing contour was manually placed inside the FAZ of the smoothed image and iteratively moved by the segmentation program toward the FAZ boundary. Five tests with different initializing curves were run on each of 26 images to assess reproducibility. The accuracy of the program was also validated by comparing results obtained by the program with the FAZ boundaries manually delineated by medical retina specialists. Interobserver performance was then evaluated by comparing delineations from two of the experts. RESULTS. One-way analysis of variance indicated that the disparities between different tests were not statistically significant, signifying excellent reproducibility for the computer program. There was a statistically significant linear correlation between the results obtained by automation and manual delineations by experts. CONCLUSIONS. This automated segmentation program can produce highly reproducible results that are comparable to those made by clinical experts. It has the potential to assist in the detection and management of foveal ischemia and to be integrated into automated grading systems. PMID:20130279

Zheng, Yalin; Gandhi, Jagdeep Singh; Stangos, Alexandros N; Campa, Claudio; Broadbent, Deborah M; Harding, Simon P

2010-07-01

118

Cholyllysyl fluroscein and related lysyl fluorescein conjugated bile acid analogues.  

PubMed Central

There have been attempts to couple bile acids to fluorescein to permit their visualization during studies of physiology and pathophysiology. Although conjugation has been achieved by many, the product differed in many respects from the parent bile acid congener. We describe lysylfluorescein conjugated bile acid analogues (LFCBAA) synthesized in our laboratory as model divalent "unipolar" molecules. We have determined LFCBAA properties including their water:octanol partition coefficient, HPLC retention time and critical micellar concentration and compared them with their parent bile acid congeners. Cholyl lysylfluorescein (CLF) and lithocholyl lysylfluoroscein (LLF) have properties similar to cholylglycine (CG) and glycolithocholate (GLC), respectively. In human and rat hepatocytes uptake of CLF follows Michaelis-Menten kinetics with K(m) and Vmax similar to CG. Biliary excretion rates of CLF and LLF closely resemble those of CG and GLC in both normal and mutant TR- rats which lack the multiorganic anion transporter (MOAT), strongly supporting the notion that CLF and LLF are substrates for the canalicular bile salt transporter (cBST). The close similarity of hepatocyte uptake and biliary secretion of these LFCBAA and their parent bile acid congeners makes them potentially useful probes for the intracellular visualization of bile salt movement and deposition in various models of bile formation and secretion.

Mills, C. O.; Milkiewicz, P.; Saraswat, V.; Elias, E.

1997-01-01

119

Influences of Acid on molecular forms of fluorescein and photoinduced electron transfer in fluorescein-dispersing sol-gel titania films.  

PubMed

Fluorescein-dispersing titania gel films were prepared by the acid-catalyzed sol-gel reaction using a titanium alkoxide solution containing fluorescein. The molecular forms of fluorescein in the films, depending on its acid-base equilibria, and the complex formation and photoinduced electron transfer process between the dye and titania surface were investigated by fluorescence and photoelectric measurements. The titanium species were coordinated to the carboxylate and phenolate-like groups of the fluorescein species. The quantum efficiencies of the fluorescence quenching and photoelectric conversion were higher upon excitation of the dianion species interacting with the titania, i.e. the dye-titania complex. This result indicated that the dianion form was the most favorable for formation of the dye-titania complex exhibiting the highest electron transfer efficiency. Using nitric acid as the catalyst, the titania surface bonded to the fluorescein instead of the adsorbed nitrate ion during the steam treatment. The dye-titania complex formation played an important role in the electron injection from the dye to the titania conduction band. PMID:24502447

Nishikiori, Hiromasa; Setiawan, Rudi Agus; Miyashita, Kyohei; Teshima, Katsuya; Fujii, Tsuneo

2014-07-01

120

Enzootic Pneumonia of Pigs: Identification of a Causative Mycoplasma in Infected Pigs and in Cultures by Immunofluorescent Staining  

PubMed Central

Immunofluorescent staining has been used to identify Mycoplasma hyopneumoniae in smears of broth cultures, in infected pig testicle cell cultures, and in frozen cut sections of pneumonic lungs from field and experimentally produced cases of enzootic pneumonia. In the pneumonic pig lung, fluorescent staining was limited to the surface of the bronchial and bronchiolar epithelium and to the contained exudate. In a series of trials using experimentally infected pigs fluorescence was not detected until 25 days post-infection and was regularly seen in pigs killed thereafter. Porcine immune globulin precipitated from the serum of experimentally infected pigs and conjugated with fluorescein isothiocyanate was reactive and specific for the detection of M. hyopneumoniae. Immune globulin conjugates prepared from the serum of hyperimmunized rabbits were reactive but in some cases produced a faint non-specific staining of frozen tissue sections. No such non-specific reactions were noted on stained culture smears or cell cultures. Fluorescence was not seen in known positive preparations stained with non-immune pig globulin conjugates or in preparations from uninoculated cell cultures or pigs, stained with non-immune or immune globulin conjugates. Mycoplasma hyorhinis was detected by immunofluorescent staining with homologous conjugates, in smears of broth cultures and in tissue sections from pigs with polyserositis. Immunofluorescent staining was found to be species specific and useful for the early species identification of mycoplasma isolated from pigs. ImagesFig. 1.Fig. 2.Fig. 3.Fig. 4.

L'Ecuyer, C.; Boulanger, P.

1970-01-01

121

Blood Parasites in Egyptian Animals. Part I. On the Staining of Blood Films with Giemsa Stains.  

National Technical Information Service (NTIS)

Proper use of Giemsa Stain is essential in the diagnosis of blood parasites from blood films. Such factors as staining time, dilution of stain, and pH of diluent markedly affect the staining quality of Giemsa reagent. Inconsistent staining resulting from ...

S. A. Michael, R. R. Maronpot, B. A. M. Botros

1970-01-01

122

Staining of Bacteria in Tissue Sections: A Reliable Gram Stain Method.  

National Technical Information Service (NTIS)

A consistently reliable Gram stain procedure which differentially stains both Gram-positive and Gram-negative bacteria in tissue sections is presented. A comparison of this new method with well known Gram stain methods demonstrates its superiority in diff...

H. C. Hopps R. C. Brown

1972-01-01

123

Activity Stain for Dihydroxy-Acid Dehydratase,  

National Technical Information Service (NTIS)

An activity stain has been devised for the dihydroxy-acid dehydratase. When applied to polyacrylamide gel electropherograms of crude soluble extracts of Escherichia coli, it detected a single electromorph. The intensity of staining increased with the amou...

C. F. Kuo T. Mashino I. Fridovich

1987-01-01

124

Fluorescein angiography findings in a case of Rubinstein-Taybi syndrome  

PubMed Central

The purpose of this report is to describe the fluorescein angiography findings in a case of Rubinstein-Taybi syndrome. Fundus photography and fluorescein angiography were performed on a 6-year-old male with Rubinstein-Taybi syndrome due to CREB binding protein gene mutation. Fundus photography showed glaucomatous cupping and diffusely attenuated retinal vasculature. Choroidal vasculature was prominent due to diffuse retinal atrophy with scattered focal retinal pigment epithelial changes. Fluorescein angiography showed retinal vascular attenuation, prolonged arteriovenous transit time with delayed venous filling, late small vessel leakage, and 360 degrees of peripheral avascularity. Peripheral retinal avascularity and retinal vascular inflammation evidenced by late small vessel leakage can be demonstrated by fluorescein angiography in the retinal dystrophy of Rubinstein-Taybi syndrome.

Jacobs, David J; Sein, Julia; Berrocal, Audina M; Grajewski, Alana L; Hodapp, Elizabeth

2012-01-01

125

Lifetime-based sensing of the hyaluronidase using fluorescein labeled hyaluronic acid  

PubMed Central

In this report we propose a lifetime-based sensing (LBS) for the detection of hyaluronidase (HA-ase). First, we heavily label hyaluronan macromolecules (HA) with fluorescein amine. The fluorescein labeled HA (HA-Fl) has a weak fluorescence and short fluorescence lifetime due to an efficient self-quenching. Upon the addition of HA-ase, the brightness and lifetime of the sample increase. The cleavage of an HA macromolecule reduces the energy migration between fluorescein molecules and the degree of the self-quenching. A first order of the cleavage reaction depends on the amount of the HA-ase enzyme. We describe an HA-ase sensing strategy based on the lifetime changes of the fluorescein labeled HA in the presence of HA-ase. We demonstrate that the calibration of the sensing response is the same for the average lifetime as for a single exponential decay approximation, which significantly simplifies the analysis of the sensing measurements.

Fudala, Rafal; Mummert, Mark E.; Gryczynski, Zygmunt; Rich, Ryan; Borejdo, Julian; Gryczynski, Ignacy

2011-01-01

126

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...

127

Comparison between Thermography and Fluorescein Test in the Detection of Incompetent Perforating Veins  

PubMed Central

Incompetent perforating veins of the legs were located by dynamic thermography at 69 sites and by the fluorescein test at 31. All the sites were explored by multiple local incisions. Incompetence of the veins was determined by demonstrating retrograde blood flow through these veins when severed. Explorations of these sites showed that thermography detected 94% and gave false-positive results in 6%, while the fluorescein test detected 16% and contributed 84% false-positive results. These findings show a highly significant difference (P <00005) in favour of the thermographic technique. Detection of 16% of incompetent perforating veins by the fluorescein test is statistically insignificant. This inaccuracy is thought to be due to inadequate dermal penetration of the ultraviolet rays. A positive fluorescein test probably indicates the presence of incompetent perforating veins but has little anatomical relationship to their actual site. Imagesp651-a

Elem, B.; Shorey, B. A.; Williams, K. Lloyd

1971-01-01

128

The combination of lactate and diacetate synergistically reduces cold growth in brain heart infusion broth across Listeria monocytogenes lineages.  

PubMed

Combinations of organic acids are often used in ready-to-eat foods to control the growth of Listeria monocytogenes during refrigerated storage. The purpose of this study was to quantitatively assess synergy between two organic acid growth inhibitors under conditions similar to those present in cold-smoked salmon, and to assess the effect of evolutionary lineage on response to those growth inhibitors. Thirteen strains of L. monocytogenes, representing lineages I and II, were grown at 7 degrees C in broth at pH 6.1 and 4.65% water-phase NaCl, which was supplemented with 2% potassium lactate, 0.14% sodium diacetate, or the combination of both at the same levels. Our data suggest that lineages adapt similarly to these inhibitors, as the only significant growth parameter difference between lineages was a minor effect (+/- 0.16 day, P = 0.0499) on lag phase (lambda). For all strains, lactate significantly extended lambda, from 2.6 +/- 0.4 to 3.8 +/- 0.5 days (P < 0.001), and lowered the maximum growth rate (mu(max)) from 0.54 +/- 0.06 to 0.49 +/- 0.04 log(CFU/ml)/day (P < 0.001), compared with the control. Diacetate was ineffective alone, but in combination with lactate, synergistically increased lambda to 6.6 +/- 1.6 days (P < 0.001) and decreased mu(max) to 0.34 +/- 0.05 log(CFU/ml)/day (P < 0.001). Monte Carlo simulations provided further evidence for synergy between diacetate and lactate by predicting signficantly slower growth to nominal endpoints for the combination of inhibitors. This study shows potassium lactate and sodium diacetate have significant synergistic effects on both lambda and mu(max) of L. monocytogenes at refrigeration temperature in broth, and justifies combining these inhibitors, at effective levels, in food product formulations. PMID:20377950

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

2010-04-01

129

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

130

Identification and Characterization of a Proton Pump on Lysosomes by Fluorescein Isothiocyanate-Dextran Fluorescence  

Microsoft Academic Search

Fluorescein isothiocyanate-conjugated dextran was introduced preferentially into hepatic lysosomes by intraperitoneal injection into rats. The pH in isolated lysosomes, measured by fluorescein fluorescence, was ≈ 5 and gradually increased in KCl (to 7.0) at 25 degrees C. In the presence of Mg2+, ATP caused acidification of lysosomes that was reversed by the protonophore carbonyl cyanide p-trifluoromethoxyphenylhydrazone. Mn2+, Co2+, and Fe2+

Shoji Ohkuma; Yoshinori Moriyama; Tatsuya Takano

1982-01-01

131

Fluorescent Sensors for Zn 2+ Based on a Fluorescein Platform: Synthesis, Properties and Intracellular Distribution  

Microsoft Academic Search

Two new fluorescent sensors for Zn2+ that utilize fluorescein as a reporting group, Zinpyr-1 and Zinpyr-2, have been synthesized and characterized. Zinpyr-1 is prepared in one step via a Mannich reaction, and Zinpyr-2 is obtained in a multistep synthesis that utilizes 4 ',5'-fluorescein dicarboxaldehyde as a key intermediate. Both Zinpyr sensors have excitation and emission wavelengths in the visible range

Shawn C. Burdette; Grant K. Walkup; Bernhard Spingler; Roger Y. Tsien; Stephen J. Lippard

2001-01-01

132

Synthesis and characterization of novel polymers bearing fluorescein units: thermal and optical properties  

Microsoft Academic Search

In this work, we report the synthesis and characterization of four novel series of copolymers bearing fluorescein moieties. Two monomers: fluorescein methacrylate and dimethacrylate were prepared and were copolymerized in the presence of four different acrylic monomers bearing oligo(ethylene glycol) segments: ethylene glycol phenyl ether acrylate, ethylene glycol phenyl ether methacrylate, poly(ethylene glycol) methyl ether acrylate (Mn?=?475?g\\/mol), and poly(ethylene glycol)

Jair A. Esquivel-Guzmn; Gerardo Zaragoza-Galn; Jess Ortz-Palacios; Ernesto Rivera

2012-01-01

133

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

134

Silver stains for protein in gels  

US Patent & Trademark Office Database

A silver stain method for polypeptides in gels comprising the sequential steps of photo-reversing the polypeptide-gel by treatment with an oxidizing agent, forming a latent stain image by treating the polypeptide-gel with a photosensitive salt, and developing the stain image by treating the polypeptide-gel with a reducing agent.

1983-09-20

135

Synthesis, structural, spectroscopic and thermal characteristics of disubstituted biphenyl derivative: Biphenyl-4,4?-diacetic acid  

NASA Astrophysics Data System (ADS)

A novel 4,4?-disubstituted biphenyl derivative featuring two acetic acid side arms symmetrically attached to a biphenyl system, that is biphenyl-4,4?-diacetic acid (H2bpda), has been successfully synthesized by means of the three-stage organic strategy. The synthesis product was characterized by elemental analysis, various spectroscopic techniques including FT-IR, Raman, 1H and 13C NMR as well as thermogravimetric and TG-FT-IR coupled measurements. The phase purity of material was verified on the basis of the X-ray powder diffraction. The studied compound crystallizes in the monoclinic P21/c space group with half of the molecule in the asymmetric unit. Structural studies indicate intermolecular Osbnd H⋯O hydrogen bonding between the carboxylic groups of the adjacent molecules of H2bpda. The occurrence of intermolecularly associated carboxylic groups can also be clearly seen in the vibrational spectra of the acid. On thermal analysis both in air and nitrogen an anhydrous compound demonstrates considerable thermal stability.

Sienkiewicz-Gromiuk, Justyna; G?uchowska, Halina; Tarasiuk, Bogdan; Mazur, Liliana; Rz?czy?ska, Zofia

2014-07-01

136

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

PubMed

A double fluorescence staining protocol was developed to facilitate computer based image analysis. Myofibers from experimentally treated (irradiated) and control growing turkey skeletal muscle were labeled with the anti-myosin antibody MF-20 and detected using fluorescein-5-isothiocyanate (FITC). Extracellular material was stained with concanavalin A (ConA)-Texas red. The cross-sectional area of the myofibers was determined by calculating the number of pixels (0.83 mu m(2)) overlying each myofiber after subtracting the ConA-Texas red image from the MF-20-FITC image for each region of interest. As expected, myofibers in the irradiated muscle were smaller (P < 0.05) than those in the non-irradiated muscle. This double fluorescence staining protocol combined with image analysis is accurate and less labor-intensive than classical procedures for determining the cross-sectional area of myofibers. PMID:9138529

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

1996-03-01

137

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

NASA Technical Reports Server (NTRS)

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

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

1996-01-01

138

Methodological factors affecting the results of staining frozen-thawed fertile and subfertile Japanese Black bull spermatozoa for acrosomal status.  

PubMed

In the present study, some methodological factors affecting the acrosomal staining of frozen-thawed Japanese Black bull spermatozoa were investigated by examining; the effect of fixation/permeabilization procedure on intact acrosome percentage after fluorescein isothiocyanate peanut agglutinin (FITC-PNA) staining, the acrosomal staining patterns by using two types of fluorescent probes FITC-PSA (Pisum Sativum Agglutinin) and FITC-PNA and the effect of staining methods, either smear or vial, on intact acrosome percentage. Then intact acrosome percentage was compared between the samples stained by thus established method and those simply fixed with glutaraldehyde (glutaraldehyde fixation method). A possibility that FITC-PNA staining or the glutaraldehyde fixation methods could detect any difference in intact acrosome percentage or acrosomal staining patterns between fertile and subfertile bulls was also examined. The results showed that (1) 4% paraformaldehyde fixation plus 1% Triton X-100 permeabilization was better than absolute ethanol alone, (2) FITC-PNA acrosomal labeling was more specific than FITC-PSA, (3) sperm suspensions should be smeared and gently processed before acrosomal staining rather than spotted onto glass slides after staining in vial in order to avoid excessive mechanical damage of the sperm acrosome, and (4) staining spermatozoa with FITC-PNA had no major advantages over examination of simply glutaraldehyde fixed sperm samples and both failed to detect any significant difference in intact acrosome percentage between the fertile and the subfertile bulls used here. The present study demonstrates important methodological considerations which need to be taken into account in order to design a reliable and reproducible protocol for the study of the acrosome. PMID:23182469

Almadaly, Essam; El-Kon, Ismail; Heleil, Bassiouni; Fattouh, El-Sayed; Mukoujima, Koushi; Ueda, Takuya; Hoshino, Youichirou; Takasu, Masaki; Murase, Tetsuma

2012-12-01

139

What is actually stained by rose bengal?  

PubMed

It has been believed that 1% rose bengal does not stain normal, healthy cells but rather stains degenerated or dead cells and mucous strands. In contrast to this conventional knowledge, we discovered that both commercial additive-containing and additive-free rose bengal solutions stained four different types of healthy cultured cells, including rabbit corneal epithelial cells. Rose bengal staining was rapid, dose dependent, predominantly nuclear, and detectable with the naked eye at concentrations as low as 0.05% and 0.025% for the commercial additive-containing or additive-free solutions, respectively, and with the fluorescence microscope at a concentration of 0.001%. It is surprising to discover that rose bengal is not a vital dye; after staining, cells actually lost vitality, as evidenced by instant morphologic changes, subsequent loss of cellular motility, cell detachment, and cell death. Such an intrinsic toxic effect was augmented by light exposure. The rose bengal staining of live as well as detergent-treated (Triton X-100) cells could be blocked by such tear components as mucin and albumin, suggesting that normally negative rose bengal staining is due to the protective function of the preocular tear film, ie, staining is not dictated by lack of cell vitality. These data indicate that rose bengal staining ensues whenever there is poor protection of surface epithelium by the preocular tear film; this represents a new interpretation for rose bengal stains seen in various ocular surface disorders. PMID:1637285

Feenstra, R P; Tseng, S C

1992-07-01

140

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

PubMed

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

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

2008-03-01

141

Improved measurements of intracellular nitric oxide in intact microvessels using 4,5-diaminofluorescein diacetate  

PubMed Central

4,5-Diaminofluorescein diacetate (DAF-2 DA) has been widely used for the measurement of nitric oxide (NO) in living cells and tissues. We previously established a method that demonstrated platelet activating factor (PAF)-induced endothelial NO production in intact venules using DAF-2 DA. In previous applications, the loading dye was removed from the extracellular space before NO measurements. However, in high permeability vessels, endothelial cells quickly released the accumulated intracellular DAF-2 after the washout, which compromises the NO measurement. The objective of this study was to investigate if the presence of DAF-2 DA during NO measurements could overcome the dye retention problem and enhance the sensitivity of NO detection. Experiments were conducted in individually perfused rat venules, and endothelial NO was measured using fluorescence imaging under basal and stimulated conditions with continuous perfusion of DAF-2 DA. Continuous dye perfusion was found to promote a relatively constant endothelial dye concentration in both normal and high permeability vessels throughout the experiment. With the use of this method, the basal and stimulated NO was quantified after endothelial DAF-2 concentrations reached a steady state. Our results showed enhanced sensitivity of detecting PAF-stimulated NO compared with a previous method. We also found that the hydrolyzed intracellular DAF-2, the precursor of DAF-2 triazole, contributed significantly to the measured fluorescence and that an appropriate subtraction of non-NO-dependent intracellular DAF-2 fluorescence is critical for the assessment of NO in living tissues. This method overcame the dye leakage problem, enhanced the sensitivity of NO detection, and improved NO quantification, demonstrating significant advantages over existing methodologies using DAF-2.

Zhou, Xueping

2011-01-01

142

Amylase levels in semen and saliva stains.  

PubMed

Amylase levels were determined for 148 semen samples and 20 saliva samples as well as for their corresponding stains. The effect of aging on the detectability of amylase activity in these stains was also investigated. The Phadebas amylase test was used for the quantitative assay of amylase. High levels of amylase in fluid saliva resulted in high levels being detected in saliva stains. Lower levels present in most seminal fluids produce little or no detectable amounts of amylase in stains. Interpretations are made as to the possible sources of amylase activity found in stains from laboratory casework based on both the amylase concentration and the elapsed time between collection and analysis. The evidential value of the presence or absence of amylase activity in casework stains is also discussed. PMID:2423634

Auvdel, M J

1986-04-01

143

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

PubMed Central

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

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

2013-01-01

144

Intravital multiphoton microscopy can model uptake and excretion of fluorescein in hepatic ischemia-reperfusion injury.  

PubMed

The liver is important in the biotransformation of various drugs, where hepatic transporters facilitate uptake and excretion. Ischemia-reperfusion (I/R) injury is a common occurrence in liver surgery, and the developing oxidative stress can lead to graft failure. We used intravital multiphoton tomography, with fluorescence lifetime imaging, to characterize metabolic damage associated with hepatic I/R injury and to model the distribution of fluorescein as a measure of liver function. In addition to measuring a significant increase in serum alanine transaminase levels, characteristic of hepatic I/R injury, a decrease in the averaged weighted lifetime of reduced nicotinamide adenine dinucleotide phosphate was observed, which can be attributed to a changed metabolic redox state of the hepatocytes. I/R injury was associated with delayed uptake and excretion of fluorescein and elevated area-under-the-curve within the hepatocytes compared to sham (i.e., untreated control) as visualized and modeled using images recorded by intravital multiphoton tomography. High-performance liquid chromatography analysis showed no differences in plasma or bile concentrations of fluorescein. Finally, altered fluorescein distribution was associated with acute changes in the expression of liver transport proteins. In summary, multiphoton intravital imaging is an effective approach to measure liver function and is more sensitive in contrasting the impact of I/R injury than measuring plasma and bile concentrations of fluorescein. PMID:23812606

Thorling, Camilla A; Liu, Xin; Burczynski, Frank J; Fletcher, Linda M; Roberts, Michael S; Sanchez, Washington Y

2013-10-01

145

Actin assessment in addition to specific immuno-fluorescence staining to demonstrate rickettsial growth in cell culture.  

PubMed

Rickettsiae are able to spread within infected cell mono-layers by modifying intra-cellular actin formations. The study analyzes whether a visualization of actin modifications in addition to specific immuno-fluorescence staining of rickettsiae might facilitate the proof of rickettsial growth in cell culture. Cell mono-layers of Vero E6 und BGM cells were infected with Rickettsia honei. Intra-cellular actin was fluorescence stained with TRITC-(tetra-methyl-5,6-isothiocyanate)-labeled phalloidin in addition to specific immuno-fluorescence staining of rickettsiae with FITC-(fluorescein-isothiocyanate)-labeled antibodies. DNA of bacteria and cells was counter-stained with DAPI (4,6-diamino-2-phenyl-indole). Cell cultures infected with Vaccinia virus were used as positive controls, cell cultures infected with Coxiella burnetii as negative controls. High concentrations of R. honei are necessary to demonstrate characteristic modifications of the intra-cellular actin. This effect is more pronounced in Vero E6 cells than in BGM cells. Actin staining with phalloidin is not suited for an early proof of rickettsial growth in cell culture but may confirm unclear findings in specific immuno-fluorescence staining in case of sufficient bacterial density. PMID:24265939

Frickmann, Hagen; Schrpfer, Elmar; Dobler, Gerhard

2013-09-01

146

Fluorescein dye intercalated layered double hydroxides for chemically stabilized photoluminescent indicators on inorganic surfaces.  

PubMed

A new photoactive thin film of layered double hydroxide (LDH) nanocrystals containing fluorescein dyes (LDH-F) has been developed by self-assembly of the LDH nanocrystals and well-controlled intercalation of the dyes in organic media. XRD results and absorption spectra confirmed the highly oriented interlayer arrangement of the dianionic form of the fluorescein dyes in the LDH interlayers, in which the dye molecules were electrostatically immobilized between the positively charged LDH layers with a monolayer packing structure. An intensity weighted average PL lifetime was estimated to be 1.45 ns and fluorescence lifetime imaging microscopy revealed that the individual LDH nanocrystals on the LDH-F film had largely similar lifetimes, which were ascribed to the uniform loading of fluorescein dyes onto the LDH matrix without photoluminescence quenching. PMID:24759944

Lee, Jong Hyeon; Jung, Duk-Young; Kim, Eunchul; Ahn, Tae Kyu

2014-06-14

147

Efficiency of staining hair with indocyanine green  

NASA Astrophysics Data System (ADS)

The efficiency of staining hair with indocyanine green (ICG) solution depending on type of hair, natural color, staining time and other parameters was investigated. Bonding ICG with hair material occurs due to interaction between ICG molecules and keratinocyte albumin. The penetration of ICG dye into hair meets with difficulties owing to surface protective layer.

Kulyabina, Tatyana V.; Kochubey, Vyacheslav I.

2005-06-01

148

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.

De Carlo, Sacha; Harris, J. Robin

2010-01-01

149

Specificity of Immunofluorescent Staining for Study of Aspergillus flavus in Soil  

PubMed Central

Fluorescein-labeled antiserum prepared with Aspergillus flavus strain CS was tested for specificity by staining fungi grown in soil in the vicinity of buried slides. All 14 strains of A. flavus fluoresced as intensely or nearly as intensely as the antigen control. Among 21 isolates of species of Aspergillus other than A. flavus, 17 reacted with moderate to low fluorescence at intensities readily distinguishable from that of A. flavus. The fluorescence of the remaining four cultures, and particularly A. sydowi, was indistinguishable from that of A. flavus. Fungi other than aspergilli were generally nonreactive. Interfering cross-reactions were encountered for one strain of Spicaria and one strain of Stemphylium; three isolates could not be evaluated because of interfering autofluorescence. An additional 22 isolates were either wholly negative or had a low order of fluorescence. Agglutination tests between each of the fungi and A. flavus CS serum revealed close agreement between agglutination titer and fluorescent-staining reaction. Unknown fungi freshly isolated from soil were checked for reaction to the A. flavus labeled antiserum; only one isolate gave a pronounced staining reaction, and that one proved to be a strain of A. flavus. In a simplified ecological model, the fluorescent-antibody technique was used to follow the development of A. flavus in mixed culture in soil with five other soil fungi. Images Fig. 1

Schmidt, E. L.; Bankole, R. O.

1965-01-01

150

Unconventional negative stains: Heavy metals are not required for negative staining  

Microsoft Academic Search

Salts of heavy metals (tungsten, uranium, and molybdenum) have long been used as negative stains for the transmission electron microscopy of protein molecules, supramolecular assemblies, and viruses, Negative staining still reveals details about protein structure only to around 25 (= 2.5 nm), most likely due to several major technical deficiencies in all traditional stain reagents. Experimental tests of unconventional

William H. Massover; Philip Marsh

1997-01-01

151

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

152

A rapid procedure for preparing fluorescein-labeled specific antibodies from whole antiserum: its use in analyzing cytoskeletal architecture  

Microsoft Academic Search

A rapid method for the direct conjugation of affinity-purified antibodies with fluorescein (termed DCAPA) is described. This procedure involves the immobilization of antibodies as antigen- antibody complexes on nitrocellulose blots, and subsequently the bound antibodies are reacted with fluorescein isothiocyanate. An enriched sample of smooth muscle tropomysin transferred to nitro- cellulose paper by the Western blotting procedure has been used

J. C. Talian; J. B. OLMSTED; R. D. GOLDMAN

1983-01-01

153

The application of fluorescein labeled serum proteins (FLSP) to the study of vascular permeability in the brain  

Microsoft Academic Search

1.Vascular permeability in normal and edematous brain tissue was studied by application of the fluorescein labeled serum proteins (FLSP) as well as by the use of free fluorescein isothiocyanate (FITC) marker.2.The electrophoretic studies demonstrated that the binding capacity of the FITC to albumin was of such degree that morphological observations made after injection of the free tracer can be considered

Igor Klatzo; Jaime Miquel; Richard Otenasek

1962-01-01

154

Oral fluorescein angiography: reassessment of its relative safety and evaluation of optimum conditions with use of capsules  

Microsoft Academic Search

Injection of fluorescein intravenously for fundal angiography is associated with a high incidence of minor adverse effects (21%) but a very low incidence of serious (life threatening) reactions (0.05%). A serious reaction may occur without warning in a patient with no history of atopy. There are no reports of oral fluorescein causing a serious reaction, and minor adverse effects are

A. P. Watson; E. S. Rosen

1990-01-01

155

Fluorescein transport properties across artificial lipid membranes, Caco-2 cell monolayers and rat jejunum.  

PubMed

Membrane transport characteristics of a paracellular permeability marker fluorescein were evaluated using artificial membrane, Caco-2 cell monolayers and rat jejunum, all mounted in side-by-side diffusion cells. Modified Ringer buffers with varied pH values were applied as incubation salines on both sides of artificial membrane, cell culture monolayers or rat jejunum. Passive transport according to pH partition theory was determined using all three permeability models. In addition to that, active transport of fluorescein in the M-S (mucosal-to-serosal) direction through rat jejunum was observed. The highest M-S P(app) values regarding the active transport through the rat jejunum were observed in incubation saline with pH 6.5. Fluorescein transport through the rat jejunum was inhibited by DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid) and alpha-CHC (alpha-cyano-4-hydroxycinnamic acid). Thus, we assume that two pH-dependent influx transporters could be involved in the fluorescein membrane transport through the intestinal (jejunal) epithelium. One is very likely an MCT (monocarboxylic acid cotransporter) isoform, inhibited by specific MCT inhibitor alpha-CHC, while the involvement of the second one with overlapping substrate/inhibitor specificities (most probably a member of the organic anion-transporting polypeptide family, inhibited at least partially by DIDS) could not be excluded. PMID:17129714

Berginc, Katja; Zakelj, Simon; Levstik, Lea; Ursic, Darko; Kristl, Albin

2007-05-01

156

Phase Conjugation by Degenerate Four Wave Mixing in Disodium Fluorescein Solution in Methanol.  

National Technical Information Service (NTIS)

Organic dyes are known to show the resonant type of nonlinear optical properties, including phase conjugation. In the present work, disodium fluorescein in methanol is used as an organic nonlinear medium for degenerate four wave mixing at 532 nm to see th...

H. Abdeldayem P. C. Sekhar P. Venkateswarlu M. C. Geroge

1989-01-01

157

Chemical and Physical Variables Affecting Fluorescein Isothiocyanate and Its Protein Conjugates.  

National Technical Information Service (NTIS)

The effect of pH, time, temperature, and buffer system on the fluorescence, absorbance, and stability of fluorescein isothiocyanate and its protein conjugates was studied. The effect of these parameters on dye-binding procedures was also studied. The fluo...

M. R. Klugerman

1965-01-01

158

Fluorescein: A Rapid, Sensitive, Nonlethal Method for Detecting Skin Ulceration in Fish  

Microsoft Academic Search

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

E. J. Noga; P. Udomkusonsri

2002-01-01

159

Fluorescein mercuric acetateA novel sensor for oral malodour detection  

Microsoft Academic Search

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

Sujatha Jayaraman; Ritu Walia; Nethaji Alagirisamy

2010-01-01

160

Fluorescein-Labeled Oligonucleotides for Real-Time PCR: Using the Inherent Quenching of Deoxyguanosine Nucleotides  

Microsoft Academic Search

Fluorescein-labeled oligonucleotide probes can be used to continuously monitor the polymerase chain reaction. Depending on the sequence, the fluorescence intensity of the probe is either increased or decreased by hybridization. The greatest effect is probe quenching by hybridization to amplicons containing deoxyguanosine nucleotides (Gs), giving a sequence-specific decrease in fluorescence as product accumulates. Quenching of the probes by Gs is

Andrew O. Crockett; Carl T. Wittwer

2001-01-01

161

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

Microsoft Academic Search

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

S S Hayreh; T A Weingeist

1980-01-01

162

Comparative fluorescein angiography of the normal sheep and goat ocular fundi.  

PubMed

Fluorescein angiography without sedative or anesthetic agents was evaluated in 20 normal goats and 20 normal sheep. All of the angiographic phases were observed using 20 mg/kg fluorescein IV in both species. Fundus fluorescein angiography results revealed wide stars of Winslow in the tapetal fundus, central or marginal flow during the first part of the arterial phase, delayed filling of the focal areas in the choroid near the optic disc that often coincided with others in the disc, and lack of evidence of the 'striate area' in the tapetal fundi. In goats, the angiographic times were 6.54+/-1.25 s for the arterial phase (TA), 7.80+/-1.37 s for the arterio-venous phase (TAV), and 14.13+/-2.01 s for the venous phase (TV). I1: 1.30+/-0.30 s (time elapsing between TA and TAV), and I2: 6.20+/-1.60 s (time elapsing between TAV and TV). In sheep, times were 9.54+/-2.18 s TA, 11.73+/-2.10 s TAV, and 20.86+/-2.74 s TV. I1: 2.04+/-0.75 s and I2: 8.98+/-2.47 s, respectively. Due to the large size of the fundic vessels in sheep and goats, fluorescein angiography of the retinal vasculature can facilitate the study of the different vascular diseases in these species. PMID:16409239

Galn, Alba; Martn-Surez, Eva M; Granados, M Mar; Gallardo, Jos M; Molleda, Jos M

2006-01-01

163

Observation of serum proteins in the edematous brain tissue by fluorescein-antibody technique  

Microsoft Academic Search

Histologic distribution of serum proteins within normal and edematous brain tissues was investigated with the fluorescein-antibody technique. Brain edema was produced in dogs with cold injury, as described byKlatzoet al. In normal brain large amounts of serum proteins were restricted to the vascular lumina and the presence of these proteins was sparse in nerve, glia and endothelial cells. On the

K. Someda; N. Kageyama

1968-01-01

164

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

165

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

166

Loading process of sugars into cabbage petiole and asparagus shoot apex cells by incubation with hypertonic sugar solutions  

Microsoft Academic Search

Summary The freezing tolerance of cabbage petioles and asparagus shoot apexes was increased by preincubation with 0.8 M sugar solutions. In cabbage petioles with an initial freezing tolerance of -3 C (temperature for 50% cell survival), as determined by both electrolyte leakage and fluorescein diacetate vital staining, the freezing tolerance was increased to -13 C by incubation with sorbitol solutions

Y. Jitsuyama; T. Suzuki; T. Harada; S. Fujikawa

2001-01-01

167

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. ...

168

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

Microsoft Academic Search

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

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

2002-01-01

169

Somatic hybrid potato plants after electrofusion of diploid Solanum tuberosum and Solanum phureja  

Microsoft Academic Search

Protoplasts from diploid S. tuberosum and diploid S. phureja were electrofused followed by selection of the heterokaryons with a micromanipulator. Visual identification of the heterokaryons was facilitated by fluorescein diacetate staining of the protoplasts from one of the parents, which was grown on herbicide containing medium to induce bleaching of the chlorophyll. In total, 840 heterokaryons showing red (chlorophyll) and

K. J. Puite; S. Roest; L. P. Pijnacker

1986-01-01

170

SYTO11 staining versus FISH staining: a comparison of two methods to stain Wolbachia pipientis in cell cultures  

PubMed Central

Aims The Aedes albopictus C7-10 cell line was infected with Wolbachia strains wRi and wAlbB to create C7-10R and C7-10B cell lines, respectively. We compared two different methods, FISH staining and SYTO11 staining, to describe these new Wolbachia infections in C7-10. Methods and Results Both staining methods were as efficient to stain Wolbachia. A formula was developed to quantify Wolbachia infection. The infection levels in C7-10B and C7-10R differed. The live stain SYTO11 was found to be useful to visualize Wolbachia in replicating host cells. Its potential cytotoxic effect at high concentration was investigated. Conclusions C7-10 supported two Wolbachia infections, constituting new tools to study Wolbachia-host interactions. The different infection levels suggest that wRi and wAlbB have different requirements for their survival in C7-10 host cell line. Observation of SYTO11 stained live cells gave new insights on Wolbachia segregation pattern during host cell mitosis. Significance and Impact of study Wolbachia-induced phenotypes in their arthropod and worm hosts could potentially be used to control pest populations. However, the mechanisms underlying these phenotypes are difficult to study due to Wolbachia's intracellular lifestyle. The Wolbachia infections in C7-10 described here could be used as in vitro models to investigate Wolbachia biology.

Venard, Claire M. -P.; Crain, Philip R.; Dobson, Stephen L.

2010-01-01

171

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

172

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

173

Silver staining of 2D electrophoresis gels.  

PubMed

Silver staining is used to detect proteins after electrophoretic separation on polyacrylamide gels. It -combines excellent sensitivity (in the low nanogram range) with the use of very simple and cheap equipment and chemicals. For its use in proteomics, two important additional features must be considered, compatibility with mass spectrometry and quantitative response. Both features are discussed in this chapter, and optimized silver staining protocols are proposed. PMID:22665294

Rabilloud, Thierry

2012-01-01

174

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

PubMed Central

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

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

2014-01-01

175

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

176

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

PubMed Central

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

Brunet, Aurelie; Aslam, Tashfeen; Bradley, Mark

2014-01-01

177

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

PubMed

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

Brunet, Aurlie; Aslam, Tashfeen; Bradley, Mark

2014-07-15

178

The staining of acidic proteins on polyacrylamide gels: enhanced sensitivity and stability of "Stains-all" staining in combination with silver nitrate.  

PubMed

A number of acidic proteins, such as those found in bone and dentin, are poorly resolved on acrylamide gels using Coomassie blue or silver nitrate staining. The cationic dye Stains-all allows visualization and identification of these proteins due to their differential staining: highly acidic proteins stain blue and intact proteoglycans stain purple, whereas less acidic proteins stain pink. However, the use of Stains-all is limited due to relatively poor staining sensitivity and lack of stability to light. A procedure which addresses these deficiencies has been developed utilizing established protocols for Stains-all staining followed by silver nitrate incubation and development. In this way, phosphoproteins such as osteopontin, bone sialoprotein, dentin phosphophoryn, and other acidic glycoproteins are visualized at higher sensitivity (greater than fivefold) and staining stability than normally achieved with just Stains-all. The protocol stains a greater variety of proteins than a combined alcian blue/silver staining procedure previously described. Utilizing the Stains-all/silver protocol, porcine bone osteopontin, a protein not visualized by standard silver staining, can be observed in amounts as little as 0.25 ng on polyacrylamide gels. Furthermore, densitometric scans demonstrate that the staining intensity is proportional to osteopontin amount and can be used for quantification over a range from 0.25 to 50 ng. PMID:9299020

Goldberg, H A; Warner, K J

1997-09-01

179

Negative staining and cryo-negative staining: applications in biology and medicine.  

PubMed

Negative staining is widely applicable to isolated viruses, protein molecules, macromolecular assemblies and fibrils, subcellular membrane fractions, liposomes and artificial membranes, synthetic DNA arrays, and also to polymer solutions and a variety of nanotechnology samples. Techniques are provided for the preparation of the necessary support films (continuous carbon and holey/perforated carbon). The range of suitable negative stains is presented, with some emphasis on the benefit of using ammonium molybdate and of negative stain-trehalose combinations. Protocols are provided for the single droplet negative staining technique (on continuous and holey carbon support films), the floating and carbon sandwich techniques in addition to the negative staining-carbon film (NS-CF) technique for randomly dispersed fragile molecules, 2D crystallization of proteins and for cleavage of cells and organelles. Immuno-negative staining and negative staining of affinity labeled complexes (e.g., biotin-streptavidin) are presented in some detail. The formation of immune complexes in solution for droplet negative staining is given, as is the use of carbon-plastic support films as an adsorption surface on which to perform immunolabeling or affinity experiments, prior to negative staining. Dynamic biological systems can be investigated by negative staining, where the time period is in excess of a few minutes, but there are possibilities to greatly reduce the time by rapid stabilization of molecular systems with uranyl acetate or tannic acid. The more recently developed cryo-negative staining procedures are also included: first, the high concentration ammonium molybdate procedure on holey carbon films and second, the carbon sandwich procedure using uranyl formate. Several electron micrographs showing examples of applications of negative staining techniques are included and the chapter is thoroughly referenced. PMID:24357366

Harris, J Robin; De Carlo, Sacha

2014-01-01

180

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

PubMed

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

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

2009-04-01

181

Artifactual Sulfation of Silver-stained Proteins  

PubMed Central

Sulfation and phosphorylation are post-translational modifications imparting an isobaric 80-Da addition on the side chain of serine, threonine, or tyrosine residues. These two post-translational modifications are often difficult to distinguish because of their similar MS fragmentation patterns. Targeted MS identification of these modifications in specific proteins commonly relies on their prior separation using gel electrophoresis and silver staining. In the present investigation, we report a potential pitfall in the interpretation of these modifications from silver-stained gels due to artifactual sulfation of serine, threonine, and tyrosine residues by sodium thiosulfate, a commonly used reagent that catalyzes the formation of metallic silver deposits onto proteins. Detailed MS analyses of gel-separated protein standards and Escherichia coli cell extracts indicated that several serine, threonine, and tyrosine residues were sulfated using silver staining protocols but not following Coomassie Blue staining. Sodium thiosulfate was identified as the reagent leading to this unexpected side reaction, and the degree of sulfation was correlated with increasing concentrations of thiosulfate up to 0.02%, which is typically used for silver staining. The significance of this artifact is discussed in the broader context of sulfation and phosphorylation site identification from in vivo and in vitro experiments.

Gharib, Marlene; Marcantonio, Maria; Lehmann, Sylvia G.; Courcelles, Mathieu; Meloche, Sylvain; Verreault, Alain; Thibault, Pierre

2009-01-01

182

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) [Livermore, CA; Pinkel, Daniel (Walnut Creek, CA) [Walnut Creek, CA

1998-01-01

183

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

Microsoft Academic Search

Chicchi, G. G., M. A. Cascieri, M. P. Graziano, T. Calahan and M. R. Tota. Fluorescein-Trp25-exendin-4, a biologically active fluorescent probe for the human Glp-1 receptor. Peptides 18(2) 319321, 1997.Exendin-4, a reptilian GLP-1 analogue, has been fluorescently labeled by covalently linking a fluorescein moiety onto the Trp residue yielding fluorescein-Trp25-exendin-4 (FLEX). FLEX is equipotent to GLP-1(736)-amide and exendin-4 as an

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

1997-01-01

184

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

185

Fluorescence properties of twenty fluorescein derivatives: lifetime, quantum yield, absorption and emission spectra.  

PubMed

The fluorescence lifetime (?f), emission quantum yield (?f), absorption and emission spectral data of 20 fluorescein derivatives were measured under the same conditions by using time-correlated single photon counting, steady state fluorescence and absorption methods to get comparable data. Based on the results, the factors and mechanism that control the fluorescence properties of the fluorescein dyes are discussed. Both ?f and ?f are remarkably dependent on the substitution on either xanthene or phenyl rings, but their ratio (?f/?f), i.e. rate constant of radiation process, is a constant value (0.20 10(9) s(-1)). The rate constant of nonradiation process, on the other hand, is varied with both the structure and the solvent used. PMID:24510430

Zhang, Xian-Fu; Zhang, Jianlong; Liu, Limin

2014-05-01

186

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

187

Characterization of sodium fluorescein dye immobilized within sol-gel matrix  

NASA Astrophysics Data System (ADS)

This paper details the absorption and fluorescence spectra of sodium fluorescein in aqueous solution and sol-gel thin films as a function of pH. Our results show that the fluorescence spectrum is dependent not only on the microenvironment surrounding the fluorophore but also the concentration the probe in the sol-gel matrix. The pH sensitive range is also shown to be a function of the emission wavelength.

Holmes-Smith, A. Sheila; Yang, Yatao; Campbell, Michael; Wallace, Peter A.

1996-12-01

188

Fundus changes in mesangiocapillary glomerulonephritis type II: clinical and fluorescein angiographic findings.  

PubMed Central

Previously we have demonstrated a deposit in Bruch's membrane in a single case of mesangiocapillary glomerulonephritis type II. We studied a group of patients with this disease and described extensive clinical and fluorescein angiographic abnormalities, which were in marked contrast to the findings in a group of patients with other forms of glomerulonephritis. This finding contributes to our understanding of the pathophysiology of the complex of the retinal pigment epithelium, Bruch's membrane, and choriocapillaris. Images

Duvall-Young, J; Short, C D; Raines, M F; Gokal, R; Lawler, W

1989-01-01

189

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.

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

2014-01-01

190

Cytoskeletal F-Actin Patterns Quantitated with Fluorescein Isothiocyanate-Phalloidin in Normal and Transformed Cells  

Microsoft Academic Search

Actin in cultured fibroblasts is organized into a complex set of fibers. Patterns of organization visualized with antibody to actin are similar but not identical to those visualized with fluorescein isothiocyanate-phalloidin (Fl-phalloidin), a chemical that binds to F-actin polymer with a dissociation constant of 2.7 10-7 M [Wulf, E., Deboben, A., Bautz, F. A., Faulstich, H. & Wieland, T. (1979)

M. Verderame; D. Alcorta; M. Egnor; K. Smith; R. Pollack

1980-01-01

191

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

Microsoft Academic Search

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

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

1993-01-01

192

Solvatochromic Studies of Fluorescein Dianion in N, N-Dimethylformamide\\/Water and Dimethylsulphoxide\\/Water Mixtures  

Microsoft Academic Search

The absorption, excitation and fluorescence spectra of the fluorescein dianion (FL) in N, N-Dimethylformamide (DMF)\\/water (H2O) and Dimethylsulphoxide (DMSO)\\/H2O solvent mixtures have been investigated. It is found that the absorption ?max and emission maxima EX, nax are both hypsochromic shifted when the H2O content in the solvent mixtures increases. However, the shoulder peaks ?s remain constant at 483. 5nm within

Ming Fat Choi; Peter Hawkins

1994-01-01

193

A new approach for visualisation of dye leakage in fluorescein angiography  

Microsoft Academic Search

INTRODUCTION: Fluorescein angiography (FA) is the more common investigation performed for macular diseases.1 Frozen FA pictures are obtained but direct visualisation of the kinetics of FA is possible only once, by the investigator. The kinetics of FA examination is imagined from static FA pictures based on our experience.23 We evaluated a new software that re-create automatic pseudo-movie from static pictures,

Giuseppe Querques; Gisele Soubrane; Eric H Souied; Nicola Delle Noci

2007-01-01

194

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

195

An evaluation of elastic tissue staining.  

PubMed

The Special Procedures Laboratory of the Department of Pathology at M.D. Anderson Hospital was concerned with the nationwide shortage of hematoxylin. The amount of this dye required for various staining solutions was calculated to determine thrifty usage. Some solutions required more hematoxylin than was available and prompted the serious investigation of existing techniques and new methods for routine laboratory use. Evaluation of various techniques resulted in a significantly successful modification of Gomori's aldehyde fuchsin technique, which was subsequently adopted as a routine procedure for elastic tissue staining in the Special Procedures Laboratory. PMID:61727

Lambert, C; Futch, H N

1976-09-01

196

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

197

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

PubMed

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

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

2014-01-01

198

[Observation of the epicerebral microcirculation studied by fluorescein angiography (author's transl)].  

PubMed

Fluorescein angiography was carried out by modification of method as described by Feindel et al. The hemisphere was widely exposed to allow visualization of surface blood vessels. In the experimental room, the dye was injected rapidly either through a fine polyethylen catheter placed in the lingual artery or thorough the cannula in the femoral vein. For "lingual" angiography 1.6 ml of 1% sodium fluorescein were used, while for "femoral" angiography, 4 ml of 10% solution were injected. In the operating room 4 ml of 1% sodium fluorescein are rapidly injected into an internal carotid catheter. Serial photographs of the passage of the dye through the surface vessels of the hemisphere were taken with a motor-drive Nikon camera at interval of 0.4 seconds, or longer when indicated, starting at the time of the injection. The shutter was synchronized with the discharge of a rapid re-charging stroboscopic light. A wratten gelatin filter 47A (Kodak) was used over the light and a Nikon filter Y52 over the camera lens for Kodak high speed ektachrome film to obtain color photographs. The timing of the interval between photographs was measured to within in 0.4 seconds by recording from a ink written oscillography synchronized with the shutter. Thus the velocity of flow in individual vessels could be calculated from the serial photographs. PMID:986017

Shibata, S; Mori, K

1976-07-01

199

Topical flurbiprofen in extracapsular cataract surgery: effect on pupillary diameter and iris fluorescein leakage.  

PubMed

Recent clinical studies indicate that flurbiprofen, a cyclooxygenase inhibitor, prevents miosis and breakdown of the blood-aqueous barrier during cataract surgery. Yet based on clinical and experimental data, some researchers do not agree that flurbiprofen prevents miosis. We conducted a double-blind clinical study of the effects of topical 0.03% flurbiprofen sodium on intraoperative pupillary diameter and iris fluorescein leakage after extracapsular cataract surgery. In the first phase of the study, 120 patients who had extracapsular cataract extraction with posterior chamber intraocular lens implantation were randomly assigned to receive preoperative topical flurbiprofen or a placebo, with or without intraoperative epinephrine, in addition to the standard regimen. In the second phase, 60 of the 120 patients continued the topical flurbiprofen or placebo for one month postoperatively. Iris fluorescein angiography was performed at the end of the first and the fourth weeks. The results indicate that flurbiprofen was significantly more effective (P < .0001) in maintaining mydriasis during surgery than the placebo. This action was enhanced by intraoperative epinephrine. Flurbiprofen also significantly reduced (P < .001) postoperative iris fluorescein leakage. PMID:8229720

Cillino, S; Casanova, F; Cucco, F; Ponte, F

1993-09-01

200

2-D Registration and 3-D Shape Inference of the Retinal Fundus from Fluorescein Images  

PubMed Central

This study presents methods to 2-D registration of retinal image sequences and 3-D shape inference from fluorescein images. The Y-feature is a robust geometric entity that is largely invariant across modalities as well as across the temporal grey level variations induced by the propagation of the dye in the vessels. We first present a Y-feature extraction method that finds a set of Y-feature candidates using local image gradient information. A gradient-based approach is then used to align an articulated model of the Y-feature to the candidates more accurately while optimizing a cost function. Using mutual information, fitted Y-features are subsequently matched across images, including colors and fluorescein angiographic frames, for registration. To reconstruct the retinal fundus in 3-D, the extracted Y-features are used to estimate the epipolar geometry with a plane-and-parallax approach. The proposed solution provides a robust estimation of the fundamental matrix suitable for plane-like surfaces, such as the retinal fundus. The mutual information criterion is used to accurately estimate the dense disparity map, while the Y-features are used to estimate the bounds of the range space. Our experimental results validate the proposed method on a set of difficult fluorescein image pairs.

Choe, Tae Eun; Medioni, Gerard; Cohen, Isaac; Walsh, Alexander C.; Sadda, SriniVas R.

2008-01-01

201

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

202

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2010 CFR

...Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and chemical solution stains for...purposes are mixtures of synthetic or natural dyes or nondye chemicals in...

2010-04-01

203

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2010 CFR

...Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and chemical solution stains for...purposes are mixtures of synthetic or natural dyes or nondye chemicals in...

2009-04-01

204

Immunofluorescence staining of paraffin sections: creating DAB staining like virtual digital images using CMYK color conversion.  

PubMed

Crystal violet treatment of formalin fixed paraffin embedded tissue slides greatly reduces the endogenous autofluorescence, and allows immunofluorescence (IF) staining with FITC or Alexa488 conjugated antibodies. Using cold CCD camera to capture the fluorescence images makes this staining method very sensitive. Here we show that combination of IF with the simultaneous recording of crystal violet induced red and Hoechst 33258 induced blue fluorescence permits the localization of the IF signal over a cytoplasmic: nuclear red:blue stain that visualizes the microscopic anatomy of the underlying tissue. To make the visual interpretation of the IF staining easier for microscopists, who are used to DAB staining over weak hematoxilin-eosin background, we created a simple color conversion procedure that turns the captured three-color fluorescence RGB (red, green, blue) images over a black background into four color CMYK (cyan, magenta, yellow, key color (black)) images. PMID:19112433

Buchynska, L; Kashuba, E; Szekely, L

2008-12-01

205

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

206

Tailoring Silver Nanodots for Intracellular Staining  

PubMed Central

Through tailored oligonucleotide scaffolds, Ag nanocluster syntheses have yielded thermally and cell culture stable silver cluster-based emitters. Optimizing ssDNA stability has enabled creation of highly concentrated and spectrally pure nanocluster emitters with strong intracellular emission. Both fixed and live-cell staining become possible, and intracellular delivery is demonstrated both through conjugation to cell penetrating peptides and via microinjection.

Choi, Sungmoon; Yu, Junhua; Patel, Sandeep A.; Tzeng, Yih-Ling; Dickson, Robert M.

2011-01-01

207

A magnetic Gram stain for bacterial detection.  

PubMed

Magnetizing: Bacteria are often classified into gram-positive and gram-negative strains by staining with crystal violet (CV). The described bioorthogonal modification of CV with trans-cyclooctene (TCO) can be used to render gram-positive bacteria magnetic with tetrazine-functionalized magnetic nanoparticles (MNP-Tz). This method allows class-specific automated magnetic detection and magnetic separation. PMID:22744868

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

2012-07-27

208

Corneal Cryopreservation Evaluated by Trypane Blue Staining  

Microsoft Academic Search

Bovine corneal tissue was employed in the study of variable factors in a method of deep-freezing, storage and thawing, using Trypane blue staining as indicator of cellular damage. Variations in cell population density in the endothelial sheet were studied as a prerequisite for the evaluation of damage during the successive stages of the freezing procedures. 90% of the cells in

Steffen Sperling

1974-01-01

209

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.

210

A method for the staining of intraosseous nerve fibers using Sihler's staining technique.  

PubMed

Understanding nerve fiber distribution in the jaw bone is important when performing invasive surgical treatments. Both microscopic and macroscopic anatomical techniques have been developed to study innervation. Conventional methods of removing and staining these structures, however, often alter structure and lack reproducibility of the resulting specimens. We sought to optimize Sihler's staining technique to stain intraosseous nerves in mandibles. Four cadaver specimens were used. The best staining of intraosseous nerve fibers was achieved by using the Plank-Rychlo solution. When the Styrene monomer was used, the resulting transparency was better than that obtained with glycerin under the same conditions. No significant differences were found between Sihler's staining procedure performed according to the conventional method and the procedure in which the second decalcification step was omitted. Our results demonstrate that applying Sihler's staining technique to bones makes them transparent and allows observation of nerves while preserving the external shape of the bone and maintaining the position of intraosseous nerve fibers. Our findings suggest our Sihler staining method for intraosseous nerve fibers can provide an intermediate resolution between macroscopic and microscopic techniques. PMID:23472877

Shiozaki, K; Miida, K; Tanaka, R; Shimoda, S

2013-08-01

211

Immunoperoxidase staining of bone marrow sections.  

PubMed

Paraffin-embedded sections of bone marrow from 131 patients were studied. The specificity of the immunoperoxidase technique for detecting intracellular kappa and lambda light chains of immunoglobulin was first evaluated in 34 cases of typical multiple myeloma or macroglobulinemia. The monoclonal light chain detected in the bone marrow myeloma was identical to the result of immunoelectrophoresis of serum or urine. By use of the same method, 97 cases in which the diagnosis was difficult were examined; cases included those of bone marrow plasmacytosis, lymphocytosis, and morphologically unusual cells. All marrow cells with monoclonal immunoglobulin detected by immunoperoxidase stain eventually proved to be a B-cell neoplastic or potentially neoplastic process. This study confirms the specificity of the immunoperoxidase stain and demonstrates its value in solving clinical diagnostic problems, that is, the diagnosis of nonsecretory myeloma and unusual morphology, differentiating the monoclonal plasmaproliferative disorders from reactive plasmacytosis or lymphoma from lymphocytosis. PMID:6794902

Hitzman, J L; Li, C Y; Kyle, R A

1981-12-01

212

Meconium staining of the brainstem with open myelomeningocele.  

PubMed

Meconium staining of open myelomeningoceles has been reported to occur both prenatally and postnatally, but meconium staining of the brainstem has not been previously documented. The authors present a case of meconium staining of the brainstem in an infant with a meconium-stained myelomeningocele, Chiari malformation Type II, and hydrocephalus and discuss possible implications for prenatal and perinatal care. PMID:23157393

Lam, Sandi; Grandhi, Ramesh; Greene, Stephanie

2013-02-01

213

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

214

Staining localization of ferric reduction on roots  

Microsoft Academic Search

Tomato plants (cv. Rutgers') were grown in nutrient solutions without and with FeEDDHA or at low phosphorus levels to evaluate the formation of prussian blue (Fe4[Fe(CN)6]3?nH2O), a stain used in determining the site of ferric reduction on roots. The effect of Fe source, Fe concentration, solution pH, and time were evaluated. Prussian blue was compared with nitro?BT, and agar systems

P. F. Bell; R. L. Chaney; J. S. Angle

1988-01-01

215

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

216

An optimized device for staining electron microscopy grids.  

PubMed

Proper staining of grids is critical for transmission electron microscopy (TEM). Staining must be done as quickly as possible using minimal reagents and with consideration for the environment. We developed a new device for efficient staining of multiple TEM grids. We studied reagent evaporation, rinsing volume, flow rate and re-use of uranyl acetate, and provide here a procedure for efficient staining using the new device. Our device permits TEM grids to be stained with less reagent than alternative staining apparatuses; staining requires a total volume of 260 ?l for five grids. Reagent evaporation is less than 6% even if used at 37 C. Moreover, our staining apparatus reduces chemical waste and shortens experiment time by staining several grids simultaneously. Our staining device is a compromise between time-consuming single grid processing and expensive commercial devices that consume large amounts of reagents. PMID:23962218

Mathieu, E; Benmlih, K; Fabre, R; Hemmerle, J

2014-01-01

217

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

PubMed

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

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

2012-10-14

218

Effects of sevoflurane, isoflurane and halotane anaesthesia on fluorescein angiographic phases of dogs: a comparative study.  

PubMed

A fluorescein angiography method was developed to compare the onset and the total duration of the fluorangiographic phases between three anaesthetic protocols in six healthy mixed-breed dogs. The animals were anaesthetized three times. Each dog received, as pre-anaesthetic protocol, atropine (10 micrograms/kg intramuscularly), and as a sedative, romifidine (80 micrograms/kg intravenously). Fifteen minutes later, induction of anaesthesia was delivered with propofol (1 mg/kg intravenously) and maintained either with sevoflurane (SEVO group), isoflurane (ISO group) or halothane (HAL group) for 30 min in all cases. Some angiographic, cardiovascular and respiratory variables were registered during the procedure. Recovery times were also registered. Angiographic variables recorded were: onset of the arterial phase (TA), onset of the arteriovenous phase (TAV), onset of the venous phase (TV), complete arterial phase duration (I1), complete arteriovenous phase duration (I2) and I1 plus I2 (I3). Mean heart rate, mean arterial pressure, systolic arterial pressure, diastolic arterial pressure, respiratory rate, tidal volume, arterial oxygen saturation and end-tidal CO2 during SEVO and ISO anaesthesia, were similar in dogs. Minute ventilation and rectal temperature were higher in dogs with SEVO than ISO. HAL produced higher arterial pressures and a lower arterial oxygen saturation than ISO and SEVO. Mean respiratory rate, rectal temperature and minute ventilation were higher in HAL. Pulse rate, end-tidal CO2 and tidal volume were similar in the dogs of the three groups. No differences in recovery times were found. The fluorescein angiographic times were within the normal range. There were no significant differences between protocols in I1, I2 or I3. HAL produced a significant increase of all temporal variables (TA, TAV and TV) when compared with ISO; TA was higher in HAL than SEVO-treated dogs. All protocols appear to be safe and effective for inducing and maintaining general anaesthesia in healthy dogs for performing fluorescein angiography. PMID:11475901

Martn, E; Redondo, J I; Molleda, J M; Santisteban, J M; Lpez, R; Gmez-Villamandos, R

2001-06-01

219

Optical fibre temperature sensor based on fluorescein and rhodamine codoped polymer layer  

NASA Astrophysics Data System (ADS)

The article presents the luminescent based optical fiber transducer. The new construction of polymer optical fibre sensor with resonant energy transfer is shown. The idea and fabrication process of low cost optode is presented. The fluorescein and rhodamine B codoped polymethylmethacrylate (PMMA) sensitive layer exhibits the wide range of absorption spectrum ensures high source to optode spectrum alignment. The luminescent response under 430 and 470nm Light Emitting Diode (LED) source is shown. The experimental characteristic of sensor in the range from 293 K to 403 K is shown. The article presents also the potential applications of presented sensor.

Miluski, Piotr; Dorosz, Dominik; Kochanowicz, Marcin; ?mojda, Jacek

2013-10-01

220

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

PubMed

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

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

2013-01-01

221

Fluorescein angiographic findings in three patients with long-term intravitreal liquid silicone.  

PubMed Central

The long-term retinal effects of intravitreal liquid silicone (ILS) remain controversial. In this study fundus fluorescein angiographic findings in three patients with long-term ILS are presented. Sluggish or absent blood flow was observed in retinal arterioles that lay in close proximity to the ILS, and the arterioles themselves appeared narrowed. It is suggested that ILS may have a long-term effect on the retinal vasculature, owing either to direct vascular damage, secondary to damage to the neuroretina, or to the ILS preventing diffusion of oxygen into the vitreous cavity. Images

Gray, R H; Cringle, S J; Constable, I J

1989-01-01

222

Control of Listeria monocytogenes in ready-to-eat meats containing sodium levulinate, sodium lactate, or a combination of sodium lactate and sodium diacetate.  

PubMed

This study investigated the use of sodium levulinate to prevent outgrowth of Listeria monocytogenes in refrigerated ready-to-eat (RTE) meat products. Turkey breast roll and bologna were formulated to contain 1%, 2%, or 3% (w/w) sodium levulinate, 2% sodium lactate, a 2% combination of sodium lactate and sodium diacetate (1.875% sodium lactate and 0.125% sodium diacetate), or no antimicrobial (control). Samples of the RTE products were sliced, inoculated with 10(2) to 10(3) CFU/cm(2) of a 5-strain cocktail of L. monocytogenes, vacuum packaged, and stored at refrigeration temperature for 0 to 12 wk. Counts reached 10(8) CFU/cm(2) on control turkey roll product after 8 wk, and over 10(7) CFU/cm(2) on control bologna after 12 wk. Addition of 2% or more sodium levulinate to turkey roll and 1% or more sodium levulinate to bologna completely prevented growth of L. monocytogenes during 12 wk of refrigerated storage. A consumer taste panel with pathogen-free samples found no differences in the overall liking among the preparations of turkey roll or among preparations of bologna. These results show that sodium levulinate is at least as effective at inhibiting outgrowth of L. monocytogenes in RTE meat products as the current industry standards of lactate or lactate and diacetate, and levulinate addition does not alter the overall liking of the RTE meat products. PMID:18577007

Thompson, R L; Carpenter, C E; Martini, S; Broadbent, J R

2008-06-01

223

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

224

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

PubMed Central

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

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

2013-01-01

225

The influence of halogen derivatives of thyronine and fluorescein on the dipole potential of phospholipid membranes.  

PubMed

The effects of halogen derivatives of thyronine (tetraiodotironine and triiodothyronine) and fluorescein (Rose Bengal, phloxine B, erythrosin, eosin Y, and fluorescein) on the dipole potential of membranes composed of diphytanoylphosphocholine, diphytanoylphosphoserine, and diphytanoylphosphoethanolamine were investigated. A quantitative description of the modifying action of the agents was presented as characteristic parameters of the Langmuir adsorption isotherm: the maximum changes in the dipole potential of the membrane at an infinitely high concentration of modifiers and the desorption constant, characterizing their inverse affinities to the lipid phase. It was shown that the iodine-containing hormones led to a less significant reduction in the dipole potential of phospholipid membranes compared to the xanthene dyes, Rose Bengal, phloxine B, and erythrosin. The latter were characterized by the highest affinity for the lipid membranes compared to tetraiodotironine and triiodothyronine. It was found that the effect of iodine-containing hormones and xanthene dyes on the membrane dipole potential was caused by their uncharged and charged forms, respectively. PMID:25024118

Efimova, Svetlana S; Schagina, Ludmila V; Ostroumova, Olga S

2014-08-01

226

Fetal outcome in meconium stained deliveries.  

PubMed

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

Mundhra, Rajlaxmi; Agarwal, Manika

2013-12-01

227

Strategic Tool Use: Pencil Box Staining  

NSDL National Science Digital Library

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

228

Romanowsky staining in cytopathology: history, advantages and limitations.  

PubMed

If the entire discipline of diagnostic cytopathology could be distilled into a single theme, it would be the Papanicolaou stain. Yet it was the Romanowsky stain upon which the discipline of cytopathology was founded. Both stains are used today in the cytopathology laboratory, each for a different and complementary purpose. We trace the history of cytopathological stains and discuss the advantages and limitations of Romanowsky-type stains for cytological evaluation. We also provide suggestions for the advantageous use of Romanowsky-type stains in cytopathology. PMID:21395493

Krafts, K P; Pambuccian, S E

2011-04-01

229

Treatment of port-wine stains: analysis  

SciTech Connect

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

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

1987-08-01

230

Disciform detachment of the macula. II. Fluorescein and indocyanine green fluorescence angiographic findings in juvenile haemorrhagic macular choroidopathy.  

PubMed

Six patients with juvenile haemorrhagic mascular choroidopathy were studied with fluorescein and indocyanine green fluorescence (ICG) angiography, and red-light and red-free light photography in different stages of the disease. The primary lesion consisted of multifocal, whitish, dot-like areas of choroidal infiltration showing hyperfluorescence in the late phase of the fluorescein angiograms. Red-light photographs revealed depigmentation of the pigment epithelium overlying the choroidal lesion, and clearly demonstrated the subsequent pigment-ring lesion. Fluorescein angiograms revealed subretinal neovascularization at the site of the disciform-stage choroidal lesion. ICG angiograms revealed the choridal lesion to be located in the region of greatest supply of short posterior ciliary arteries, wheras the lesion itself remained underfilled throughout the angiogram suggesting vascular decompensation at the site of the lesion. The results suggest a vascular basis, namely intravascular coagulation in the central choriocapillaris, for this uveitis entity. PMID:577366

Saari, M

1977-06-01

231

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

232

Delivery room management of meconium-stained infant.  

PubMed

This article discusses the historical background, epidemiology, and pathophysiology of meconium-stained amniotic fluid and provides current concepts in delivery room management of meconium-stained neonate including the current Neonatal Resuscitation Program guidelines. PMID:23164180

Bhat, Rama; Vidyasagar, Dharmapuri

2012-12-01

233

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.

234

Staining protocols for human pancreatic islets.  

PubMed

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

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

2012-01-01

235

Basophil Counting With a New Staining Method Using Alcian Blue  

Microsoft Academic Search

Difficulties in obtaining reproducible and accurate enumeration of circulating baso- phils with existing techniques have ham- pored investigation of this infrequent cell population. A new basophil staining method is described that employs alcian blue dye for staining of heparin within basophils at low pH and in the presence of lanthanum ions. Basophil recognition is facilitated by reducing nonspecific nuclear staining.

Harriet S. Gilbert; Leonard Ornstein

1975-01-01

236

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

237

Staining methods applied to glycol methacrylate embedded tissue sections  

Microsoft Academic Search

The use of glycol methacrylate (GMA) avoids some technical artifacts, which are usually observed in paraffin-embedded sections, providing good morphological resolution. On the other hand, weak staining have been mentioned during the use of different methods in plastic sections. In the present study, changes in the histological staining procedures have been assayed during the use of staining and histochemical methods

P. S Cerri; E Sasso-Cerri

2003-01-01

238

Flash fluorometer made from off-the-shelf photographic equipment to measure tissue levels of fluorescein.  

PubMed

A device to measure fluorescein in tissue has been constructed from standard photographic equipment--an electronic strobe and a flashmeter both covered with interference filters. The instrument works well in the light and need not touch the area being measured, an advantage over existing fluorometers. The instrument has been used to measure the amount of dye in flaps in rats, pigs, and three humans. The results revealed that the amount of dye in a freshly made flap was rarely as much as in normal skin, and skin with less than 20 percent of the dye of control areas usually sloughed, although there were exceptions. In the future the instrument will be improved, and its readings will be compared to those obtained from radioactive microspheres, the present "gold standard" of techniques to measure vascularity. The instrument can be used to estimate the blood supply to any tissue and seems to be as reliable as the dermofluorometers already on the market. PMID:2909065

Meyers, B; Valencia, S

1989-01-01

239

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

240

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

NASA Astrophysics Data System (ADS)

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

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

2013-12-01

241

[Use of the dia-contact method in ophthalmology. A new method of fluorescein angiography presentation].  

PubMed

An innovative presentation of fluorescein angiograms on radiographic film, called "dia-contact", is described. Photographs are produced by the classical procedure of contact-printing on subtraction film and are of very high quality resolution. Less expensive than conventional prints, the resultant document is easy to copy and file. The reading of this document with the help of two + 10 lenses, allows a sequential analysis of the angiogram and a stereoscopic interpretation alternatively. The image can be enlarged without loss of resolution by means of a microfilm reader device. This method requires a certain amount of self discipline, in that a new habit must be learned. However it rapidly proved itself invaluable in the author's daily practice. PMID:6674320

Mouillon, M; Gilliotte, J; Romanet, J P

1983-01-01

242

Validation of an automated fluorescein method for determining bromide in water  

USGS Publications Warehouse

Surface, atmospheric precipitation and deionized water samples were spiked with ??g l-1 concentrations of bromide, and the solutions stored in polyethylene and polytetrafluoroethylene bottles. Bromide was determined periodically for 30 days. Automated fluorescein and ion chromatography methods were used to determine bromide in these prepared samples. Analysis of the data by the paired t-test indicates that the two methods are not significantly different at a probability of 95% for samples containing from 0.015 to 0.5 mg l-1 of bromide. The correlation coefficient for the same sets of paired data is 0.9987. Recovery data, except for the surface water samples to which 0.005 mg l-1 of bromide was added, range from 89 to 112%. There appears to be no loss of bromide from solution in either type of container.Surface, atmospheric precipitation and deionized water samples were spiked with mu g l** minus **1 concentrations of bromide, and the solutions stored in polyethylene and polytetrafluoroethylene bottles. Bromide was determined periodically for 30 days. Automated fluorescein and ion chromatography methods were used to determine bromide in these prepared samples. Analysis of the data by the paired t-test indicates that the two methods are not significantly different at a probability of 95% for samples containing from 0. 015 to 0. 5 mg l** minus **1 of bromide. The correlation coefficient for the same sets of paired data is 0. 9987. Recovery data, except for the surface water samples to which 0. 005 mg l** minus **1 of bromide was added, range from 89 to 112%. Refs.

Fishman, M. J.; Schroder, L. J.; Friedman, L. C.

1985-01-01

243

Vascular and avascular retinae in mammals. A funduscopic and fluorescein angiographic study.  

PubMed

Intraretinal blood vessels are present in some and absent in other vertebrate species, including the mammals. Among the marsupials, both vascular and avascular retinae are seen. We determined the funduscopic appearance of the eye, investigated the functional aspects of ocular blood flow in both types of retina in marsupials and compared our results with known patterns in placental mammals. The Australian polyprotodont marsupials, the Tasmanian devil, Sarcophilus harrisii, and the quoll, Dasyurus viverrinus, together with an American polyprotodont, the Virginia opossum, Didelphis virginiana, demonstrate variable degrees of tapetal differentiation, pigmentation and a very close parallel course of their intraretinal arteries and veins over considerable distances. Using the technique of fluorescein angiography, we found that retinal blood flow in the 3 vascular Australian species commenced with arterial filling. Early venous was seen next, followed by the capillary blush. This unusual sequence of vascular flow differs from that of the arterial-capillary-venous filling seen in placental mammals. This difference is most likely a consequence of the known looped, end artery organisation found within marsupial nervous systems, of which the retinae are a part. The 2 diprotodont marsupials examined, the brushtail possum, Trichosurus vulpecula, and the sugar glider, Petaurus breviceps, possess avascular retinae. Only a small residual tuft of fluorescein-impermeable vessels projects from the optic disc into the vitreous. Interestingly, the structural complexity of the central visual system in diprotodonts all of whom possess avascular retinae) is commonly accepted as being greater than that of the stem polyprotodont line (which possess vascular retinae). If retinal function matches this internal complexity, then retinal avascularity may, as in birds, be associated with superior vision. However, as the retinae of these mammals clearly lack any nutritive mechanisms directly analogous to those in the retinae of, say, birds or the megachiropteran bats, their retinal nutritive pathways remain enigmatic. PMID:2375974

Buttery, R G; Haight, J R; Bell, K

1990-01-01

244

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.

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

1995-01-01

245

Comments on the history of the Biological Stain Commission, Inc.  

PubMed

Nearly 89 years ago, the Society of American Bacteriologists appointed Dr. Harold Conn to form a committee to standardize the stains and dyes used in biological and medical research and diagnosis. Dr. Conn's efforts led to formation of the Committee on the Standardization of Biological Stains, later incorporated as the Biological Stain Commission. This article traces some of the events and factors that shaped the course of the Biological Stain Commission into its current form and functions. Its principal function is to ensure that the biological and medical communities have access to high quality, dependable and consistent biological dyes and stains. PMID:21838612

Penney, D P

2012-01-01

246

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

247

Adsorption characteristics of arsenic(III) and arsenic(V) on iron(III)-loaded chelating resin having lysine-N ?,N ?-diacetic acid moiety  

Microsoft Academic Search

An iron(III)-loaded chelating resin (Fe-LDA) with lysine-N?,N?-diacetic acid functional groups has been prepared and its adsorption characteristics for arsenic(III) and arsenic(V) have been examined. Arsenic(V) was strongly adsorbed to the resin in the pH range from 2 to 4, while arsenic(III) was moderately adsorbed between pH 8 and 10. The isotherm data for arsenic(V) at pH 3.5 fitted well to

H. Matsunaga; T. Yokoyama; R. J. Eldridge; B. A. Bolto

1996-01-01

248

Protein gel staining methods: an introduction and overview.  

PubMed

Laboratory scientists who encounter protein biochemistry in many of its myriad forms must often ask: is my protein pure? The most frequent response: run a denaturing SDS polyacrylamide gel. Running this gel raises another series of considerations regarding detection, quantitation, and characterization and so the next questions invariably center on suitable protein gel staining and detection methods. A total protein profile can be determined with the colorimetric methods embodied in Coomassie Blue and silver staining methods, or increasingly, with fluorescent stains. Protein quantitation can be done following staining, with fluorescence- and instrumentation-based methods offering the greatest sensitivity and linear dynamic range. Protein posttranslational modifications such as phosphorylation and glycosylation can be reliably determined with several fluorescence-based protocols. Staining and detection with two or more different stains can be done in series to establish relative profiles of modified versus total protein or to assess purity at two levels of quantitative sensitivity. The choice of staining method and protocol depends on the required rigor of detection and quantitation combined with available instrumentation and documentation capabilities. Other considerations for staining methods include intended downstream analytical procedures such as mass spectrometry or peptide sequencing, which preclude some methods. Nonfixative staining methods allow western blotting after gel staining. Laboratory custom and budget or intellectual curiosity may be the ultimate determinate of the chosen gel staining protocol. PMID:19892191

Steinberg, Thomas H

2009-01-01

249

Non-contact ultra-widefield retinal imaging and fundus fluorescein angiography of an infant with incontinentia pigmenti without sedation in an ophthalmic office setting.  

PubMed

When fluorescein angioscopy or angiography is required in an infant, it is usually performed in the operating theater or neonatal unit. We report a case of an infant with incontinentia pigmenti in whom we were able to acquire angiographic information in an office setting by using an ultra-widefield non-contact system with oral fluorescein. PMID:23607978

Patel, Chetan K; Fung, Timothy H M; Muqit, Mahiul M K; Mordant, David J; Geh, Vernon

2013-06-01

250

Cell-trappable quinoline-derivatized fluoresceins for selective and reversible biological Zn(II) detection.  

PubMed

The synthesis and spectroscopic characterization of two new, cell-trappable fluorescent probes for Zn(II) are presented. These probes, 2-(4,5-bis(((6-(2-ethoxy-2-oxoethoxy)quinolin-8-yl)amino)methyl)-6-hydroxy-3-oxo-3H-8 xanthen-9-yl)benzoic acid (QZ2E) and 2,2'-((8,8'-(((9-(2-carboxyphenyl)-6-hydroxy-3-oxo-3H-xanthene-4,5-diyl)bis(methylene))bis(azanediyl))bis(quinoline-8,6-diyl))bis(oxy))diacetic acid (QZ2A), are poorly emissive in the off-state but exhibit dramatic increases in fluorescence upon Zn(II) binding (120 10-fold for QZ2E, 30 7-fold for QZ2A). This binding is selective for Zn(II) over other biologically relevant metal cations, toxic heavy metals, and most first-row transition metals and is of appropriate affinity (K(d1)(QZ2E) = 150 100 ?M, K(d2)(QZ2E) = 3.5 0.1 mM, K(d1)(QZ2A) = 220 30 ?M, K(d2)(QZ2A) = 160 80 ?M, K(d3)(QZ2A) = 9 6 ?M) to reversibly bind Zn(II) at physiological levels. In live cells, QZ2E localizes to the Gogli apparatus where it can detect Zn(II). It is cell-membrane-permeable until cleavage of its ester groups by intracellular esterases produces QZ2A, a negatively charged acid form that cannot cross the cell membrane. PMID:20849126

McQuade, Lindsey E; Lippard, Stephen J

2010-10-18

251

Fluorescent staining of ? opioid receptors using naltrexamine derivatives and phycoerythrin  

Microsoft Academic Search

An immunofluorescent technique that is more sensitive than radioligand binding was developed in order to detect opioid receptors expressed on leukocytes. The current study was designed to optimize the method for fluorescently labeling ? opioid receptors. For these experiments, the opioid antagonist naltrexamine was conjugated to either fluorescein (FITC-NTXamine) or biotin (biotin-NTXamine). One-step, two-step, and three-step protocols were compared to

Diane M. P Lawrence; Ian Hutchinson; Ahmad Seyed-Mozaffari; Sydney Archer; Jean M Bidlack

1997-01-01

252

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

253

Semiquantitation of bacteria in sputum gram stains.  

PubMed Central

In many clinical laboratories, bacteria seen in Gram-stained sputum smears are reported semiquantitatively, using a three- or four-category scale consisting of ratings such as numerous, moderate, rare, and none seen. The consistency with which these categories are assigned was evaluated by repeatedly presenting coded smears to seven experienced microbiology technologists. Technologists rated the same smear twice, pairs of smears prepared from the same specimen, and smears prepared after specimen refrigeration. Agreement was assessed with the weighted kappa test. Semiquantitation of gram-negative rods, gram-positive diplococci, and gram-positive cocci in clusters all showed poor reproducibility (kappa = 0.32, 0.34, and 0.17, respectively). Twenty-four percent of paired ratings differed by two or more categories. Lack of reproducibility was due mainly to the inability of the technologists to render a consistent rating when viewing the same slide on separate occasions (P less than 0.001). Variation in the rating styles of different technologists, differences between smears prepared from the same specimen, and specimen refrigeration tended to further decrease the consistency of ratings, but the reductions were not statistically significant. The quantity of potentially pathogenic bacteria in sputum smears is not estimated consistently with standard microscopy procedures and should not be reported.

Valenstein, P N

1988-01-01

254

Automatic analysis of immunocytochemically stained tissue samples.  

PubMed

An automatic colour image segmentation and cell counting software system has been developed for immunocytochemical analysis of stained tissue samples. The system was designed to count the total number of positive and negative cells in tissue samples treated with cytokine DNA probes from pigs naturally parasitised with Taenia solium metacestodes, using in situ hybridisation. A reaction index was calculated as the ratio of the number of cells with a positive reaction to the total number of cells (positives plus negatives) for each of five different probes. The objectives of automatic counting were to improve the reproducibility of the analysis and reduce the processing time of large image batches. A fast KNN classifier was used for colour segmentation. Watershed segmentation combined with edge detection was used to isolate individual cells that were then automatically labelled, using the results of the corresponding colour segmented image. Validation was performed on 122 non-training digital images with a total of 1069 positive cells and 1459 negative cells, with the following results: a mean true positive rate of 90.2% for positive cells and a mean true positive rate of 85.4% for negative cells. The corresponding mean false positive rates were 9.6% and 6.6%. The mean reaction index error of the automatic analysis was 5.35%. The processing of each digital image took 10 s on a Pentium IV PC. PMID:16411641

Armbula Coso, F; Mrquez Flores, J A; Padilla Castaeda, M A; Solano, S; Tato, P

2005-09-01

255

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.

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

2011-01-01

256

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

257

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

258

Manual hematoxylin and eosin staining of mouse tissue sections.  

PubMed

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

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

2014-01-01

259

[Study on the histochemical staining of boric acid].  

PubMed

The detection of boric acid in the tissue is of significance in investigating its toxicity. Because of this, we have devised a histochemical staining method to detect the presence of boric acid. The outline of this method follows. Frozen 12-14 microns sections, cut by a cryostat, are fixed in anhydrous ethanol and stained for 20 minutes in a protonated curcumin solution. Washing in acetic acid follows, and a red stain results if boric acid is present. This method causes a reaction, in which rosocyanin is formed by the reaction of boric acid and the protonated curcumin, and this principle is now used when an analysis of boric acid is needed. As to procedure, a 1 N concentration of sodium hydroxide is dropped onto a part of the stain to be tested, and the presence of rosocyanin is confirmed if the stain turns blue. Consequently, this staining confirms the presence of boric acid. PMID:1725792

Yoshida, M; Tokiyasu, T; Watabiki, T; Ueda, M; Ishida, N

1991-12-01

260

Factors involved in differential giemsa-staining of sister chromatids  

Microsoft Academic Search

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

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

1978-01-01

261

A review of amyloid staining: methods and artifacts.  

PubMed

Amyloid detection is very precise at this time, because several methods are available to the pathologist. Awareness of potential nonspecific staining, possible pitfalls and methods for improving the detection process are basic to enhancing the staining of amyloid and interpreting this staining. The role of the pathologist has progressed through history from the basic detection of amyloid as a substance to immunophenotypical classification of the particular amyloid present. PMID:20429750

Fernandez-Flores, A

2011-10-01

262

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

263

Diagnostic dilemma in female genital tuberculosis- staining techniques revisited.  

PubMed

Background: Tuberculosis (TB) is an increasing public health concern worldwide. On a global scale it has a devastating impact in developing nations. Genital TB, an extrapulmonary form, is not uncommon particularly in areas where pulmonary TB is prevalent. Genital TB may be asymptomatic or may even masquerade as other gynaecological conditions; hence, diagnosis requires a high degree of suspicion and the use of appropriate investigations. Objective: This study attempted to identify endometrial TB in endometrial biopsies taken from women evaluated for infertility by comparison of various staining techniques. Materials and Methods: A comparative cross sectional study was conducted from February 2011 to April 2011 in Guru Teg Bahadur Hospital, New Delhi. Endometrial biopsy specimens from 55 endometrial TB suspects were stained for acid fast bacilli by Ziehl Neelson staining and Gabbet staining. The biopsy samples were also subjected to Auramine Phenol fluroscent staining and H and E staining. Culture on Lowenstein Jensen medium was taken as the gold standard. Results: Three samples were culture positive giving positivity rate of 5.4%. Considering culture as the gold standard the senstivities of ZN, Gabbet, fluorescent and H and E staining were 33, 33, 66, and 66% respectively while their specificities were 100, 100, 98, and100% respectively. Conclusion: Combination of fluorescent staining techniques along with one of the acid fast staining techniques or histopathology achieves sufficient sensitivity and specificity for the diagnosis of female genital tuberculosis. There is an urgent need for developing definitive diagnostic methods to make a conclusive diagnosis of genital TB. PMID:24639789

Kashyap, Bineeta; Srivastava, Namita; R Kaur, Iqbal; Jhamb, Rajat; K Singh, Deepak

2013-07-01

264

Groundwater dating using radiocarbon in fulvic acid in groundwater containing fluorescein  

NASA Astrophysics Data System (ADS)

Natural DO14C is recognized as one of the most useful tracers in the estimation of groundwater age. Fluorescent dye is commonly used as an indicator of drilling fluid contamination during borehole investigation. Fluorescein (FS) is one of the most frequently used fluorescent dyes, yet as it contains little radiocarbon it may affect DO14C age when it is mixed with natural DOC. In this study, fulvic acid (FA) was isolated from groundwater containing FS and DO14C value of isolated FA was measured. Separation methods were proposed by using the difference between sorption and desorption behavior of FA and FS onto synthetic adsorbent resin. DO14C measurement on FA from a mixture of FA and FS is estimated by removing FS from the mixture and correcting for the amount of C derived from FS. Furthermore, DO14C age is compared with the groundwater age estimated by He. The results show that the values of DO14C age for separated FA estimated by the two methods had good agreement with those that corresponded to groundwater age estimated by He. This result indicates that DO14C is a useful indicator of groundwater age even for groundwater contaminated with FS.

Nakata, Kotaro; Kodama, Hiroki; Hasegawa, Takuma; Hama, Katsuhiro; Iwatsuki, Teruki; Miyajima, Tohru

2013-05-01

265

In-vivo pharmacokinetic study of two fluorescein derivatives by fluorescence spectroscopy  

NASA Astrophysics Data System (ADS)

We have already demonstrated the ability of fluorescence spectroscopy and imaging to measure the pH of superficial tissues using pH sensitive fluorescent probes. The purpose of this study was to investigate the in vivo behavior of such fluorescent probes. We report the monitoring of tissue fluorescence after injection of two fluorescein derivatives (carboxyfluorescein and biscarboxyethyl-carboxyfluorescein). The in vivo study was performed on anaesthetized adult Wistar rats. After laparotomy, CF or BCECF solution was injected into the penial vein. Fluorescence spectra were recorded during one hour using an optical multichannel analyzer coupled to a CCD camera. Fiber optic was placed alternatively on the liver area or on the skin. Blood samples were collected and fluorescence was measured in vitro. A clear linear relationship between dose and fluorescence intensity was found in liver for these fluorescent markers. Concerning spectral characteristics, it was found that CF and BCECF spectra show a shift compared to in vivo maximum emission peak and BCECF emission peak was different when recorded in the liver and in the skin. Differences of kinetic profiles are also observed between CF and BCECF. The BCECF derivative displays a fluorescence peak in the liver two minutes after injection, while CF fluorescence peak is observed seven minutes after injection. Clearance of skin fluorescence is slower than the plasmatic one indicating that dye elimination in superficial blood vessels does not follow the same pharmacokinetic behavior. Based on these preliminary findings, fluorescence spectroscopy appears as a tool in pharmacokinetic study in situ and in vivo.

Soulie-Begu, Sylvie; Devoisselle, Jean-Marie; Mordon, Serge R.

1995-12-01

266

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 ?EST 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

267

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

PubMed Central

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

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

2000-01-01

268

Comparison of observed and predicted normalized air concentrations for 56-m releases of fluorescein particles  

SciTech Connect

Centerline ground-level normalized air concentration measurements made at Hanford, Washington, for the short-term release of fluorescein particles from a height of 56 m were compared with a Gaussian plume atmospheric dispersion model using two different sets of dispersion parameters and two different methods of classifying atmospheric stability. The ratio of the predicted air concentration to the observed air concentration is strongly dependent on the downwind distance being considered. All four methods have a tendency to underpredict near the source, sometimes by many orders of magnitude, and to overpredict at the farthest distances considered (12.8 km). Such a tendency must be taken into account when assessing the impact (on man) of short-term pollutant releases to the atmosphere. In general, the results of this study highlight the difficulty of choosing a set of dispersion parameters and estimating atmospheric stability for use in assessment activities. These results also show, however, that such specification is important in determining the accuracy of Gaussian plume atmospheric dispersion model predictions.

Miller, C.W.; Little, C.A.; Cotter, S.J.

1980-01-01

269

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

270

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.

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

2003-01-01

271

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 1.2610-9 (, 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

272

Volatilization of fluorescein mercuric acetate by marine bacterial from Minamata Bay  

SciTech Connect

Some bacteria that live in a mercury-polluted environment are resistant to mercury compounds. A majority of these mercury-resistant bacterial have been found to volatilize organic as well as inorganic mercury compounds into elemental mercury vapor by means of their enzymes. One compound, fluorescein mercuric acetate (FMA) has long been in use as a disinfectant in hospitals; yet, there has been little definitive information on bacterial resistance to this compound. Minamata Bay has been heavily polluted by mercury, which has caused methylmercury poisoning in humans, called Minamata disease. Sediments from the Bay still contain high concentrations of mercury. The percentage of mercury-resistant bacteria in the total bacterial count is higher in these sediments than in those of other marine environments. FMA-pollution, however, has not been reported. Research into the mechanism of bacterial resistance to FMA will not only add to our general understanding of the ability of certain bacteria to resist mercury, but will also help in defining the role bacteria play in the mercury cycle of a mercury-polluted environment. The purpose of the present study is to determine the mechanism of resistance to FMA of the FMA-resistant bacteria living in the Bay.

Nakamura, Kunihiko (National Institute for Minamata Disease, Minamata City Kumamoto (Japan))

1989-05-01

273

Phase conjugation by degenerate four wave mixing in disodium fluorescein solution in methanol  

NASA Technical Reports Server (NTRS)

Organic dyes are known to show the resonant type of nonlinear optical properties, including phase conjugation. In the present work, disodium fluorescein in methanol is used as an organic nonlinear medium for degenerate four wave mixing at 532 nm to see the intensity dependence of the phase conjugate signal at different concentrations of the solution. It is observed that the maximum reflectivity of the signal occurs in a concentration range of 5 x 10(exp -3)/cu cm to 1.2 x 10(exp -2) g/cu cm. It is also observed that the intensity of the signal drops suddenly to less than half of its maximum outside the concentration range mentioned above. An investigation of the phase conjugate signal intensity by changing the delay time between probe signal and the forward pump is also examined. Briefly discussed is the possibility of population grating in dye liquids as a source of enhancing the third order susceptibility besides the other techniques mentioned in reference. The experiment is done by beam splitting the second harmonic (532 nm) of Nd:YAG laser, Q-switched at 20 pulses/sec (pulse width is approximately 8 and 200 mJ per pulse).

Abdeldayem, Hossin; Sekhar, P. Chandra; Venkateswarlu, P.; Geroge, M. C.

1989-01-01

274

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

PubMed

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

2014-04-01

275

Fluorescein-methotrexate transport in rat choroid plexus analyzed using confocal microscopy.  

PubMed

One function of the vertebrate choroid plexus (CP) is removal of potentially toxic metabolites and xenobiotics from cerebrospinal fluid (CSF) to blood for subsequent excretion in urine and bile. We have used confocal microscopy and quantitative image analysis to follow transport of the large organic anion fluorescein-methotrexate (FL-MTX) from bath (CSF side) to blood vessels in intact rat CP and found concentrative transport from CSF to blood. With 2 microM FL-MTX in the bath, steady-state fluorescence in the subepithelium and vascular spaces exceeded bath levels by 5- to 10-fold, but fluorescence in epithelial cells was below bath levels. FL-MTX accumulation in subepithelium and vascular spaces was reduced by NaCN, Na removal, and by other organic anions, e.g., MTX, probenecid, and estrone sulfate. Increasing medium K 10-fold had no effect. None of these treatments affected cellular accumulation. However, two observations indicated that apical FL-MTX uptake was indeed mediated: first, cellular accumulation was a saturable function of medium substrate concentration; and second, digoxin and MK-571 reduced FL-MTX accumulation in the subepithelial/vascular spaces but also increased cellular accumulation severalfold. In the presence of digoxin and MK-571, cellular accumulation was concentrative, specific, and Na dependent. Thus transepithelial FL-MTX transport involved the following two mediated steps: Na-dependent uptake at the apical membrane and electroneutral efflux at the basolateral membrane, possibly on Oatp2 and Mrp1. PMID:15126245

Breen, Christopher M; Sykes, Destiny B; Baehr, Carsten; Fricker, Gert; Miller, David S

2004-09-01

276

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

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

1988-01-01

277

An Image-Processing Strategy for the Segmentation and Quantification of Microaneurysms in Fluorescein Angiograms of the Ocular Fundus  

Microsoft Academic Search

Digital image-processing techniques can provide an objective and highly repeatable way of quantifying retinal pathology. This study describes an image-processing strategy which detects and quantifies microaneurysms present in digitized fluorescein angiograms. After preprocessing stages, a bilinear top-hat transformation and matched filtering are employed to provide an initial segmentation of the images. Thresholding this processed image results in a binary image

Timothy Spencer; John A. Olson; Kenneth C. McHardy; Peter F. Sharp; John V. Forrester

1996-01-01

278

Pregnancy-Related changes in the mouse oviduct and uterus revealed by differential binding of fluoresceinated lectins  

Microsoft Academic Search

The binding of 20 fluorescein isothiocyanate (FITC)-labeled lectins to various portions of the pregnant and non-pregnant murine oviduct and uterus was studied by fluorescence microscopy. Five lectins (from Ricinus communis (RCA-I), Maclura pomifera (MPA), Triticum vulgare (wheat germ-WGA), Bauhinia purpurea (BPA), and Ulex europeus (UEA-I)) reacted differentially with the epithelium of pregnant as compared with the non-pregnant uterus. The binding

M.-C. Lee; T.-C. Wu; Y.-J. Wan; I. Damjanov

1983-01-01

279

Multiple binding sites of fluorescein isothiocyanate moieties on myoglobin: photophysical heterogeneity as revealed by ground- and excited-state spectroscopy  

Microsoft Academic Search

Fluorescein isothiocyanate (FITC)myoglobin conjugates were synthesized with a binding stoichiometry of one to three fluorophores per protein. FITC binding sites were determined by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS). Five lysine residues and the N-terminal amino group were identified as preferential binding sites. The ground and excited-state absorption spectra and the fluorescence decay of the conjugates in the

Gisela Grunwaldt; Sophie Haebel; Christian Spitz; Martin Steup; Ralf Menzel

2002-01-01

280

On the luminescence of luminol in DMSO in the presence of potassium superoxide-18-crown-6-ether and fluorescein  

Microsoft Academic Search

Luminol solution in DMSO in the presence of [18C6K]+ O2? supramolecular complex (achieved from KO2 and 18-Crown-6 (18C6)-ether) is chemiluminescent, and its intensity depends on the complex concentration. Using fluorescein (Fl) as an energy acceptor in this system, the luminescence energy transfer process from chemically excited species, aminophtalate dianion, to Fl could be evidenced. On the basis of Frster theory,

Mariana Voicescu; Marilena Vasilescu; Titus Constantinescu; Aurelia Meghea

2002-01-01

281

Validation of the reconstituted high-density lipoprotein (rHDL) drug delivery platform using dilauryl fluorescein (DLF)  

Microsoft Academic Search

Dilauryl fluorescein (DLF) is a lipid soluble molecule that becomes fluorescent when lauric acid is removed by hydrolysis\\u000a The purpose of these studies was to evaluate DLF as a potential probe for the function of reconstituted high-density lipoproteins\\u000a (rHDL) as hydrophobic drug transport vehicles. The DLF containing rHDL nanoparticles were characterized regarding their physical\\/chemical\\u000a properties, including molecular diameter, molecular weight,

Walter J. McConathy; Sulabha Paranjape; Linda Mooberry; Sabitha Buttreddy; Maya Nair; Andras G. Lacko

2011-01-01

282

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

Microsoft Academic Search

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

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

1999-01-01

283

Virus-Specific Responses of Heterosigma akashiwo to Infection  

Microsoft Academic Search

We used flow cytometry to examine the process of cell death in the bloom-forming alga Heterosigma akashiwo during infection by a double-stranded DNA virus (OIs1) and a single-stranded RNA virus (H. akashiwo RNA virus (HaRNAV)). These viruses were isolated from the same geographic area and infect the same strain of H. akashiwo. By use of the live\\/dead stains fluorescein diacetate

Janice E. Lawrence; Corina P. D. Brussaard; Curtis A. Suttle

2006-01-01

284

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

285

Islet-like cell clusters: viability, cell types, and subretinal transplantation in pancreatectomized cats  

Microsoft Academic Search

Summary We investigated a method for isolating sufficient feline islets of Langerhans to restore normoglycaemia following transplantation into the subretinal space of pancreatectomized cats. Collagenase digestate of feline pancreas was maintained in serum-free tissue culture medium for 1-9 days. Viability of islet-like cell clusters (ICC) was assessed with ethidium bromide and fluorescein diacetate staining; cell types were identified immunohistochemi- cally.

Takatoshi Maeno; Makoto Inoue; Sherif N Embabi; Daijiro Miki; Diane L Hatchell

2006-01-01

286

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

287

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

288

Solute concentration-dependent contact angle hysteresis and evaporation stains.  

PubMed

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

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

2014-07-01

289

Methylene blue selectively stains intestinal metaplasia in Barrett's esophagus  

Microsoft Academic Search

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

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

1996-01-01

290

Beyond bacteria: Interpreting fungal elements in the Gram stain  

Microsoft Academic Search

Many clinical specimens sent for routine culture and susceptibility testing require direct examination with a Gram-stained smear. The technologist in the routine microbiology section will examine the direct smear for bacteria and yeast but may miss the presence of other pathogens, such as fungal elements (FE). In our experience, FE present in direct Gram-stained smears have remained undetected on a

Subhash K Mohan

2004-01-01

291

Digital staining of pathological tissue specimens using spectral transmittance  

Microsoft Academic Search

Staining of tissue specimens is a classical procedure in pathological diagnosis to enhance the contrast between tissue components such that identification and classification of these components can be easily performed. In this paper, a framework for digital staining of pathological specimens using the information derived from the L-band spectral transmittance of various pathological tissue components is introduced, particularly the transformation

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

2005-01-01

292

Silver-stained structures in mammalian meiotic prophase  

Microsoft Academic Search

Silver staining of mammalian spermatocytes revealed, in light microscopy, synaptonemal complex and structures within the sex vesicle. It is feasible to follow the chromosome pairing phenomenon from zygotene to pachytene by examining the behavior of synaptonemal complexes. Nucleolus organizer regions take heavy silver stain in pachytene but are no longer detectable in later stages of meiosis.

S. Pathak; T. C. Hsu

1979-01-01

293

Chlorination effect on the fluorescence of nucleic acid staining dyes  

Microsoft Academic Search

An alternative to culture methods for the control of drinking water disinfection would use fluorescent dyes that could evidence the nucleic acid damages provoked by sodium hypochlorite treatment. The two dyes selected in this study, SYBR Green II RNA gel stain and TOTO-1 iodide, efficiently stain nucleic acids (DNA and RNA) and quite poorly the other biomolecules considered (Bovine serum

M. H. Phe; M. Dossot; J. C. Block

2004-01-01

294

Glycogen; its ultrastructural staining characteristics and distribution in some nematodes  

Microsoft Academic Search

In view of erratic staining of glycogen in thin sections of nematode tissues, a study of cytochemical staining using the periodic acid-thiosemicarbazidesilver protein technique along with diastase control reactions was carried out on the nematode Capillaria hepatica. Glycogen is readily demonstrated by this technique and its morphology and distribution has been examined in six nematode species. This cytochemical technique is

Kenneth A. Wright; Terry A. Dick

1972-01-01

295

An experimental method for testing novel retinal vital stains  

Microsoft Academic Search

There is uncertainty surrounding the safety of the vital stains currently used to assist macular surgery, and there may be other agents that are more suitable. This study aimed to validate a method of screening retinal vital stains for their potential surgical utility. Bovine retina was exposed to test agents at a range of concentrations. Masked observers determined the minimum

Timothy L. Jackson; Lewis Griffin; Brendan Vote; Jost Hillenkamp; John Marshall

2005-01-01

296

Comparison of sulfur hexafluoride, fluorescein and rhodamine dyes and the bacteriophage PRD-1 in tracing subsurface flow  

NASA Astrophysics Data System (ADS)

We compared velocities of the subsurface flow from a mounded onsite septic system towards a depressional wetland with three types of tracer; an inert gas, sulfur hexafluoride (SF 6), two fluorescent dyes, fluorescein and rhodamine WT, and a viral tracer, the bacteriophage PRD-1. The movement of both fluorescent dyes was significantly retarded in the soils compared to both SF 6 and PRD-1. In experiments using injection solutions containing both a dye and SF 6, fluorescein was found to move at least 3-4 times slower than SF 6, and rhodamine was not observed away from the drainfield. In contrast, the velocities calculated from SF 6 data are very similar to the velocities calculated from the PRD-1 data obtained during the same experiment. At a second site, the movement of fluorescein was half as fast and not as extensive as the movement of SF 6. The results of these experiments indicate that fluorescent dyes may underestimate velocities of effluent from septic systems adjacent to seasonal wetlands. In contrast, SF 6 was found to perform similarly to the viral tracer PRD-1.

Harden, Harmon S.; Chanton, Jeffrey P.; Rose, Joan B.; John, David E.; Hooks, Mark E.

2003-06-01

297

Control of Listeria monocytogenes on commercially-produced frankfurters prepared with and without potassium lactate and sodium diacetate and surface treated with lauric arginate using the Sprayed Lethality in Container (SLIC(R)) delivery method.  

PubMed

Viability of Listeriamonocytogenes was monitored on frankfurters formulated with or without potassium lactate and sodium diacetate at a ratio of ca. 7:1 and treated with lauric arginate (LAE; 22 or 44ppm) using the Sprayed Lethality in Container (SLIC(R)) delivery method. Without antimicrobials, pathogen numbers remained relatively constant at ca. 3.3logCFU/package for ca. 30d, but then increased to ca. 8.4logCFU/package over 120d. Regardless of whether or not lactate and diacetate were included, when treated with LAE, pathogen numbers decreased from ca. 3.3logCFU/package to ca. 1.5logCFU/package within 2h, but then increased to 7.3 and 6.7logCFU/package, respectively, after 120d. When frankfurters were formulated with lactate and diacetate and treated with LAE, pathogen numbers decreased by ca. 2.0logCFU/package within 2h and remained relatively unchanged over the 120d. These data confirm that LAE provides an initial lethality towards L. monocytogenes and when used in combination with reduced levels/ratio of lactate and diacetate as an ingredient for frankfurters provides inhibition throughout shelf life. PMID:20374905

Porto-Fett, A C S; Campano, S G; Smith, J L; Oser, A; Shoyer, B; Call, J E; Luchansky, J B

2010-06-01

298

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

PubMed

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

Pirie, C G; Alario, A

2014-03-01

299

In situ measurement of fluorescein release by collagen shields in human eyes.  

PubMed

Fluorescein (F) and FITC Dextran (FD; MW 4400) have been used as inert analogs of active drugs to examine the factors controlling the rate of loss of drugs from collagen shields in vivo. The diffusion constants in the shield (120 microns thick) determined from in vitro release experiments were found to be 0.42 x 10(-6) for F and 0.16 x 10(-6) cm2/sec for FD at 34 degrees C and these values correspond to 0.7 and 0.27 min-1 rates of loss for the exponential parts of their release. In the eyes of 6 subjects, the F loaded shields lost the dye at an average rate of 0.016 min-1 initially, increasing to 0.026 min-1 at the end of an hour. Somewhat higher values were noted with FD and were attributed to variations of manufacturing of the shields. Comparing the rates of loss in in vitro and in vivo, it is clear that the latter is not controlled by the rate of diffusion in the shield, but is limited by the rates of tear secretion and flow over the surfaces of the shield. The penetration of F into the anterior chamber from a series of four drops applied 12 min apart was compared with that from a shield soaked in the drop solution to determine the enhancement in the bioavailability. This showed that the shield increased the penetration into the eye by about 16 times over that from a single drop. However, the rate of loss of F from the shield in vivo is only about 8 times slower than that from a drop instilled in the eye.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7518372

Srinivas, S P

1994-04-01

300

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; Tting, Thomas

2014-06-01

301

Transmission and small-angle scattering of light by nematic liquid crystal-cellulose diacetate composite with self-organized structure  

NASA Astrophysics Data System (ADS)

Electrooptical properties of thin films of nematic liquid crystal-cellulose diacetate (NLC-CD) composite with self-organized structure. The composite samples were prepared at different rates of solvent evaporation on substrates inclined at various angles. The best optical contrast is obtained in composite films formed on substrates inclined at 45. The dependence of the small-angle scattering of light on the control electric field strength in the NLC-CD composite differs from the analogous dependence known for the traditional polymer-dispersed liquid crystal composites. It is established that the classical theory of the small-angle scattering does not adequately describe structural characteristics of the NLC-CD composite with self-organized structure.

Sadovoy, A. V.; Shipovskaya, A. B.

2010-12-01

302

Effect of Beta-Cyclodextrin on the Fluorescence, Absorption and Lasing of Rhodamine 6G, Rhodamine B and Fluorescein Disodium Salt in Aqueous Solutions.  

National Technical Information Service (NTIS)

The fluorescence, absorption and lasing of three xanthene dyes, rhodamine 6G, rhodamine B and the disodium salt of fluorescein were examined in aqueous solutions with and without added Beta-cyclodextrin. Beta-cyclodextrin enhances both fluorescence and la...

I. R. Politzer K. T. Crago T. Hampton J. Joseph J. H. Boyer

1989-01-01

303

Sodium diacetate and sodium lactate affect microbiology and sensory and objective characteristics of a restructured turkey breast product formulated with a fibrin cold-set binding system.  

PubMed

Research was conducted to manufacture and evaluate a restructured turkey breast product using the Fibrimex cold-set binding system, sodium diacetate (NaD), and sodium lactate (NaL) and to ascertain effects of the treatments on proximate composition, pH, psychrotrophic organisms, water activity, onset of rancidity (TBA), thaw loss, cooking yields, and objective color, and sensory characteristics. Whole turkey breasts were cut into 5-cm-thick strips; treated with either water only (control), 1.5% NaL, 2.0% NaL, 0.1% NaD, 1.5% NaL + 0.1% NaD, or 2.0% NaL + 0.1% NaD; blended with Fibrimex ingredients; stuffed into casings; and stored at -30 degrees C for 0, 1, 2, and 3 mo. After each storage period, frozen chubs were tempered at 4 degrees C, sliced into 1-cm-thick steaks, packaged in retail trays, stored at 0 degrees C to simulate retail storage, and analyzed after 0, 2, 4, 6, 8, and 10 d. Sodium diacetate used alone or in combination with NaL reduced (P < 0.05) growth of psychrotrophic organisms and had no adverse effects on water activity, pH, cooking yield, fat, moisture, protein, objective color, onset of rancidity, and sensory characteristics (juiciness, turkey flavor intensity, and tenderness). Panelists reported slight off-flavor in all steaks treated with NaL. Treating steaks with NaL alone or in combination with NaD resulted in increased (P < 0.05) ash content. Sodium lactate also functioned to minimize thaw loss in the frozen restructured turkey product. PMID:20181879

Mohammed Shafit, H; Williams, S K

2010-03-01

304

DNA comet Giemsa staining for conventional bright-field microscopy.  

PubMed

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

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

2014-01-01

305

DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy  

PubMed Central

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

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

2014-01-01

306

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.

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

2013-01-01

307

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

PubMed

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

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

2013-09-01

308

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

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

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

309

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

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

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

310

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

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

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

311

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

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

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

312

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

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

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

313

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

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

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

314

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

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

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

315

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

316

Prevention of Sap Stain and Mold in Packaged Lumber.  

National Technical Information Service (NTIS)

In the maintenance of wood quality during shipment and storage, protection against sap stain and mold in unseasoned, sodium tetrachloro-phenate treated lumber stored in commercial-size packages held in Vancouver, Canada, and in Princes Risborough, England...

J. W. Roff A. J. Cserjesi G. W. Swann

1974-01-01

317

25. FIRST AND SECOND FLOOR, STAINED GLASS WINDOW REMOVED SOUTHEAST ...  

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

25. FIRST AND SECOND FLOOR, STAINED GLASS WINDOW REMOVED SOUTHEAST SIDE AT LANDING BETWEEN FIRST AND SECOND FLOOR, PHOTOGRAPHED AT STORAGE LOCATION - Masonic Temple, 1111-1119 Eleventh Street, Altoona, Blair County, PA

318

24. FIRST AND SECOND FLOOR, STAINED GLASS WINDOW REMOVED FROM ...  

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

24. FIRST AND SECOND FLOOR, STAINED GLASS WINDOW REMOVED FROM SOUTHEAST SIDE AT LANDING BETWEEN FIRST AND SECOND FLOOR, PHOTOGRAPHED AT STORAGE LOCATION - Masonic Temple, 1111-1119 Eleventh Street, Altoona, Blair County, PA

319

45. FIRST AND SECOND FLOOR, STAINED GLASS WINDOW REMOVED FROM ...  

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

45. FIRST AND SECOND FLOOR, STAINED GLASS WINDOW REMOVED FROM SOUTHEAST SIDE AT LANDING BETWEEN FIRST AND SECOND FLOOR, PHOTOGRAPHED AT STORAGE LOCATION - Masonic Temple, 1111-1119 Eleventh Street, Altoona, Blair County, PA

320

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

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

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

321

Staining of extracellular polymeric substances and cells in bioaggregates  

Microsoft Academic Search

Multiple fluorochrome experiments with as many fluorochromes as possible are desired for exploring the detailed structure\\u000a of bioaggregates. Spectral peak interference and other practical limitations, however, restrict the maximum number of stains\\u000a used simultaneously to three. This current study proposes a sixfold labelled scheme to stain the total cells, dead cells,\\u000a proteins, lipids, and ?- and ?-polysaccharides in bioaggregates. Two

Ming-Yuan Chen; Duu-Jong Lee; Joo-Hwa Tay; Kuan-Yeow Show

2007-01-01

322

News from the Biological Stain Commission no. 15.  

PubMed

In the 15(th) issue of News from the Biological Stain Commission (BSC), under the heading of Regulatory affairs, the Biological Stain Commission's International Affairs Committee presents information from the plenary meetings of the International Standards Organization ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on August 22-24, 2012 in Berlin, Germany. An additional discussion of the use of food dyes in India also is included. PMID:24646106

Lyon, H O; Horobin, R W

2014-04-01

323

Histological identification of Helicobacter pylori: comparison of staining methods  

PubMed Central

AimTo determine whether two recently described staining methods (the modified McMullen's and the Helicobacter pylori silver stain HpSS methods) used for the histological identification of H pylori organisms are superior to two established techniques (the modified Giemsa and anti-H pylori antibody immunostain) in terms of availability, reproducibility, rapidity, sensitivity, and cost. MethodsHistological sections from 63 paired gastric biopsies from adult patients previously investigated for dyspepsia were stained with the four methods and these were assessed blindly and independently by two observers. Of the 63 patients, 30 were originally negative in all tests for H pylori infection, 30 were positive, and the remaining three cases had discordant results using a combination of five tests (rapid biopsy urease test, urea breath test, culture, serology, and histology). ResultsInterobserver agreement was best with the antibody method (98%), followed by the McMullen's (90%), Giemsa (87%), and HpSS (85%). Of the 60 "gold standard" positive and negative cases, 30 were positive by the modified Giemsa stain, 29 by the McMullen's method, 29 by HpSS, and 30 by the antibody stain. However, there were two false positives with the HpSS method. The modified Giemsa is the cheapest and easiest to perform technically. ConclusionsWhen H pylori are present, careful examination will almost always reveal them, whichever of these stains is used. However, the modified Giemsa stain is the method of choice because it is sensitive, cheap, easy to perform, and reproducible. Key Words: Helicobacter organisms histological identification staining methods

Rotimi, O; Cairns, A; Gray, S; Moayyedi, P; Dixon, M

2000-01-01

324

Methods for staining amyloid in tissues: a review.  

PubMed

The traditional way of identifying amyloid in tissue sections has been staining with Congo red and demonstration of green birefringence under crossed polarizers. The original method of Congo red staining, described by Bennhold in 1922, has undergone several modifications to improve its sensitivity, specificity, and reliability. The most common modification is the alkaline Congo red method described by Puchtler and co-workers in 1962. Specificity is improved by using freshly prepared stain and a staining solution fully saturated with sodium chloride. Amyloid proteins can be further distinguished by autoclaving or by treating the tissue with potassium permanganate or alkaline guanidine. Autoclaving the tissues at 120 C for 30 min causes protein AA to lose its affinity for Congo red. Prolongation of autoclaving to 120 min abolishes the Congophilia of protein AL, but prealbumin-related amyloid shows little or no change. Treatment of the tissue with potassium permanganate causes protein AA and B2-microglobulin amyloid to lose their affinity to Congo red. Protein AA fails to stain with Congo red after treatment with alkaline guanidine for 1 min and protein AL and systemic senile amyloid protein (SSA) after 2 hr. Familial amyloid protein (FAP), prealbumin type, can stand 2 hr of alkaline guanidine treatment without losing its ability to stain with Congo red. Other methods of detection of amyloid include fluorescent stains, e.g., thioflavin T or S, and metachromatic stains such as crystal violet. Immunofluorescence and immunoperoxidase methods are used to identify and classify amyloid proteins in tissues. Antibodies against the P component, proteins AA and AL and FAP have been used with great precision. Due to cross-reactivity, these methods do not differentiate between some types of familial and senile systemic amyloidosis. PMID:2464206

Elghetany, M T; Saleem, A

1988-07-01

325

Micrometric measurement of the density of stained odontoblast processes.  

PubMed

The embryological, structural and functional unit of the dentine-pulp complex shares the odontoblast, located in the border of the dentine pulp, with basal nuclei and organelles. The odontoblast process emerges from its apical pole. It is formed by microtubules, microfilaments and vesicles covered by membranes penetrating the dentinal tubules, isolated from the inter-tubular matrix, along the extent of the dentine. The objective of this study was to evaluate the efficacy of three staining techniques: hematoxylin-eosin, periodic acid-Schiff and Schmorl, by staining the process, from beginning to end, and compare the results with the erosion technique. Thirty human teeth were employed in the trial; after their extraction the pulp was fixated, the pieces demineralized in nitric acid at 8%, the collagen filaments eliminated with Type II Collagenase, the tissue was stained, and the measurements were made. The portions with no pulp were prepared with the erosion technique. Results: Comparing the best results obtained by staining with the values obtained with the erosion technique, the former showed lower values. Conclusion: Staining techniques show lower density of the staining processes compared with the dentinal tubules in the erosion technique. PMID:22128590

Kohli, Alicia; Pezzotto, Stella M; Garcia, Graciela; Poletto, Leonor C

2011-08-01

326

Intra-Arterial Fluorescence Angiography with Injection of Fluorescein Sodium from the Superficial Temporal Artery during Aneurysm Surgery: Technical Notes.  

PubMed

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

Ichikawa, Tsuyoshi; Suzuki, Kyouichi; Watanabe, Yoichi

2014-06-17

327

Fluorescein analogs inhibit SecA ATPase: the first sub-uM inhibitor of bacterial protein translocation  

PubMed Central

SecA is a central component of the general secretion system that is essential for bacterial growth and thus an ideal target for the development of antimicrobial agents. A series of fluorescein analogs were first screened against the ATPase activity using the truncated unregulated SecA catalytic domain. Rose Bengal (RB) and Erythrosin B (EB) were found to be potent inhibitors with IC50 values of 0.5 M and 2 M, respectively. RB and EB inhibit the catalytic SecA ATPase more than the F1F0-proton ATPase. We used three assays to test the effect of these compounds on full length SecA ATPase: in solution (intrinsic ATPase), in membrane preparation, and translocation ATPase. RB and EB show the following trend in terms of IC50 values: translocation ATPase < membrane ATPase < intrinsic ATPase. Very importantly, the potency of these fluorescein analogs in inhibiting the truncated SecA ATPase correlates with their ability to inhibit the biologically relevant protein translocation activity of SecA. The in vitro translocation of proOmpA precursors into membrane vesicles is strongly inhibited by RB with IC50 of about 0.25 M, making RB the most potent inhibitor of SecA ATPases and SecA-dependent protein translocation thus far. The ability of these compounds to inhibit SecA directly translates into antibacterial effects as well. Our findings show the value of fluorescein analogs as probes for mechanistic studies of SecA functions, and for the potential development of new antimicrobial agents with SecA as the target.

Huang, Ying-Ju; Wang, Hongyun; Gao, Fen-Biao; Li, Minyong; Yang, Hsiuchin; Wang, Binghe; Tai, Phang C.

2012-01-01

328

Crystallographic analysis of an "anticalin" with tailored specificity for fluorescein reveals high structural plasticity of the lipocalin loop region.  

PubMed

The artificial lipocalin FluA with novel specificity toward fluorescein was derived via combinatorial engineering from the bilin-binding protein, BBP by exchange of 16 amino acids in the ligand pocket. Here, we describe the crystal structure of FluA at 2.0 A resolution in the space group P2(1) with two protein-ligand complexes in the asymmetric unit. In both molecules, the characteristic beta-barrel architecture with the attached alpha-helix is well preserved. In contrast, the four loops at one end of the beta-barrel that form the entrance to the binding site exhibit large conformational deviations from the wild-type protein, which can be attributed to the sidechain replacements. Specificity for the new ligand is furnished by hydrophobic packing, charged sidechain environment, and hydrogen bonds with its hydroxyl groups. Unexpectedly, fluorescein is bound in a much deeper cavity than biliverdin IX(gamma) in the natural lipocalin. Triggered by the substituted residues, unmutated sidechains at the bottom of the binding site adopt conformations that are quite different from those observed in the BBP, illustrating that not only the loop region but also the hydrophobic interior of the beta-barrel can be reshaped for molecular recognition. Particularly, Trp 129 participates in a tight stacking interaction with the xanthenolone moiety, which may explain the ultrafast electron transfer that occurs on light excitation of the bound fluorescein. These structural findings support our concept of using lipocalins as a scaffold for the engineering of so-called "anticalins" directed against prescribed targets as an alternative to recombinant antibody fragments. PMID:12945055

Korndrfer, Ingo P; Beste, Gerald; Skerra, Arne

2003-10-01

329

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

330

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

331

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

332

Fluorescein aldehyde with disulfide functionality as a fluorescence turn-on probe for cysteine and homocysteine in HEPES buffer.  

PubMed

We developed a fluorescein aldehyde probe with disulfide functionality for the fluorescence detection of biologically important thiols. The probe displayed highly selective responses to cysteine (Cys) and homocysteine (Hcy) over glutathione (GSH) due to the rapid ring formation reaction of Cys and Hcy with the aldehyde group of the probe and the concomitant cleavage of the disulfide group followed by subsequent intramolecular cyclization. The fluorescent probe also exhibited a highly sensitive fluorescence turn-on response to Hcy with a detection limit of 2.4 ?M Hcy in HEPES buffer. PMID:23797423

Lee, Heejin; Kim, Hae-Jo

2013-08-14

333

Use of thermography and fluorescein angiography in the management of a Chilean flamingo with avascular necrosis of the wing.  

PubMed

A Chilean flamingo (Phoenicopterus chilensis) was presented to the veterinary clinic at the North Carolina Zoological Park for evaluation of acute weakness of the right wing. Results of a physical examination revealed a lack of a palpable pulse in the radial artery, which suggested occlusion or obstruction of the vessel. Radiography, thermography, and fluorescein angiography confirmed right wing injury and vascular compromise. Based on the poor prognosis for return to function associated with irreversible vascular damage, the wing was amputated. After a period of observation and treatment, the bird was returned to public exhibit. PMID:23409438

Hurley-Sanders, Jennifer L; Bowman, Karl F; Wolfe, Barbara A; Nutter, Felicia B; Sladky, Kurt K; Stoskopf, Michael K

2012-12-01

334

Analysis of surface saccharides in Trichomonas vaginalis strains with various pathogenicity levels by fluorescein-conjugated plant lectins  

Microsoft Academic Search

Certain surface saccharides of organisms from clone-derived cultures of fiveTrichomonas vaginalis strains, JH30A-cl. 1, JH31A-cl. 1, JH32A-cl. 1, JH34A-cl. 1, JH162A-cl 1, and JH384A-cl. 2, which differed in their pathogenicity for women and experimental hosts, were compared with the aid of fluorescein-conjugated plant lectins using a quantitative fluorescence method. The lectins used were: concanavalin A (Con-A), wheat germ agglutinin (WGA),

A. Wartofi; B. M. Honigberg

1983-01-01

335

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

336

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-Grnwald-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-Grnwald-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.

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

2014-01-01

337

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.

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

2013-01-01

338

A Fluorescein Tracer Release Experiment in the Hydrothermally Active Crater of Vailulu'u Volcano, Samoa  

NASA Astrophysics Data System (ADS)

Vailulu'u (Rockne) volcano marks the active end of the Samoa hotspot chain. The volcano is 4400 meters high, with a summit crater 2000 meters wide by 400 meters deep and summit peaks reaching to within 600 meters of the sea surface. The crater is hydrothermally active, as witnessed by intense particulate concentrations in the water column (values to 1.4 NTU's), a particulate smog ``halo'' surrounding the summit and extending out many kilometers, high Mn concentrations and 3He/4He ratios (values to 3.8 ppb and 8.6 Ra, respectively), and bottom-water temperature anomalies of 0.5oC. Basalts from the crater have been dated in the range 5-50 years, and likely reflect eruptions associated with a 1995 earthquake swarm. On April 3, 2001, we released a 20 kg point-source charge of fluorescein dye 30 meters above the 975m deep crater floor. The dye was dissolved in a 180 liter mixture of propanol and water, adjusted to a density 1.3 per mil heavier than the ambient water at the release depth. Released from a rubberized bag by means of a galvanic link. First detection of the released dye was 39 hours after the deployment; the dye was in a 50 meter thick layer, with a concentration peak at 900 meters (relative to the release depth of 945m). Tracking was carried out by a CTD-based fluorometer operated in tow-yo mode from the U.S.C.G. Icebreaker Polar Sea. The detection limit was 25 picograms/gram, and the maximum detected concentration was 18,000 pg/g (if evenly dispersed in the lower 150 meters of water in the crater, the expected concentration would be approx. 130 pg/g). While the dye pool was only surveyed for 4 days due to ship-transit constraints, significant horizontal and vertical dispersion was apparent. Vertical dispersion velocities were typically 0.05 cm/sec; horizontal velocities were typically higher by a factor of 10. An approximate diapycnal or eddy diffusivity, K, can be calculated from the rate of vertical spreading of the dye layer: K = Z2/2(t-t0), where Z is the layer thickness at 61 percent of the peak concentration. Our data constrain K to be in the range 200-400 cm2/sec. This may be compared with a value of 0.3 cm2/sec measured at 300m in the open ocean and a value of 5-10 cm2/sec measured in the abyssal ocean near rough topography (Ledwell et al. 1998; 2000). Clearly the water in Vailulu'u crater is in active circulation, undoubtedly driven by hydrothermal inputs. Other physical characteristics attest to this as well - gradients in potential density are small below 850 m depth in the crater, with changes ranging from 0-80 parts per billion per meter; commonly the changes in density occur in staircase fashion, and occasionally the gradients are negative. The approximate thermal output of the crater can be estimated as follows. From analysis of water samples, the total Mn budget below 800 meters is 810 kg. With an eddy diffusivity of 300 cm2/sec, the crater will lose about 66 kg of Mn per day. The typical Mn output of a 5 megawatt hot smoker on a ridge is 28 kg/day. Thus it would take several hot smokers, or a thermal output of 10 megawatts, to maintain the observed Mn budget in the crater. We believe this would make John Edmond smile: the serendipitous exploration of an active submarine volcano, in tropical waters, using an icebreaker as a ship-of-opportunity, followed by post-cruise decompression in Tisa's Bare Foot bar, Pago Pago.

Hart, S. R.; Staudigel, H.; Workman, R.; Koppers, A.; Girard, A.

2001-12-01

339

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

340

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

341

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) [Livermore, CA; Pinkel, Daniel (Walnut Creek, CA) [Walnut Creek, CA

2000-01-01

342

Transport of sodium fluorescein between the mucosal and muscular layers in man, possibly indicating functional serially-coupled exchange vessels.  

PubMed

In sixteen patients subjected to intestinal surgery, the transport of sodium fluorescein (Na-F) was measured between the mucosal and serosal-muscular layers. Experiments were carried out in intestinal anastomosis by fluorescein flowmetry (FF). The blood-flow index of the mucosal layer was about twice the serosal-muscularis, per unit tissue volume. The temporal changes in fluorescence pattern from the two layers showed that more than 90% of Na-F, eliminated from the mucosal layer, was transported to the serosal-muscular layers. In six patients, in whom Na-F was instilled intraluminarily, fluorescence was seen on the outside of the intestine after 3 min. However, when the circulation was stopped before Na-F was instilled, no fluorescence was seen, this suggests that the transport was not only due to diffusion but also to convection. The results suggest that functional serially-coupled exchange vessels may exist within the intestinal wall, and that Na-F is transported both by convection and diffusion. PMID:4092416

Perbeck, L; Nyberg, B; Thulin, L; Tydn, G

1985-12-01

343

Evolution of the silver and gold stains in neurohistology.  

PubMed

Scientific investigations depend on the reliability of the observations that can be made. This reliability is determined in part by the understanding of the techniques and technology used to make the observations. The limitations and the strengths of the methodology and the equipment used must be evaluated thoroughly. The extent to which this is and has been the case for the use of the metal based stains in neuroscience is the subject of this paper. I evaluate the metallic stains used for neuroscience from several perspectives. I review briefly the state of neurohistology prior to its "golden years," 1870-1910. Then I trace the development of the silver based stains used for neurohistology. I wanted to discuss the reasoning used by the originators of the silver based techniques in developing their specific procedures, but discovered that while procedures may be published, the methods and ideas used to arrive at the final procedures are not usually described in published work. PMID:16720522

Heinz, Tr

2005-01-01

344

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

345

New farnesane-type sesquiterpenes, hedychiols A and B 8,9-diacetate, and inhibitors of degranulation in RBL-2H3 cells from the rhizome of Hedychium coronarium.  

PubMed

Two new farnesane-type sesquiterpenes, hedychiols A and B 8,9-diacetate, were isolated from the methanolic extract of the fresh rhizome of Hedychium coronarium KOEN. cultivated in Japan. Their stereostructures were elucidated on the basis of chemical and physicochemical evidence. The inhibitory effects of isolated constituents on the release of beta-hexosaminidase from RBL-2H3 cells were examined, and hedychilactone A and coronarin D were found to show the inhibitory activity. PMID:12192135

Morikawa, Toshio; Matsuda, Hisashi; Sakamoto, Yasuko; Ueda, Kazuho; Yoshikawa, Masayuki

2002-08-01

346

The role of the Giemsa stain in cytogenetics.  

PubMed

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

Dolan, M

2011-04-01

347

Staining methods applied to glycol methacrylate embedded tissue sections.  

PubMed

The use of glycol methacrylate (GMA) avoids some technical artifacts, which are usually observed in paraffin-embedded sections, providing good morphological resolution. On the other hand, weak staining have been mentioned during the use of different methods in plastic sections. In the present study, changes in the histological staining procedures have been assayed during the use of staining and histochemical methods in different GMA-embedded tissues. Samples of tongue, submandibular and sublingual glands, cartilage, portions of respiratory tract and nervous ganglion were fixed in 4% formaldehyde and embedded in glycol methacrylate. The sections of tongue and nervous ganglion were stained by H&E. Picrosirius, Toluidine Blue and Sudan Black B methods were applied, respectively, for identification of collagen fibers in submandibular gland, sulfated glycosaminoglycans in cartilage (metachromasia) and myelin lipids in nervous ganglion. Periodic Acid-Schiff (PAS) method was used for detection of glycoconjugates in submandibular gland and cartilage while AB/PAS combined methods were applied for detection of mucins in the respiratory tract. In addition, a combination of Alcian Blue (AB) and Picrosirius methods was also assayed in the sublingual gland sections. The GMA-embedded tissue sections showed an optimal morphological integrity and were favorable to the staining methods employed in the present study. In the sections of tongue and nervous ganglion, a good contrast of basophilic and acidophilic structures was obtained by H&E. An intense eosinophilia was observed either in the striated muscle fibers or in the myelin sheaths in which the lipids were preserved and revealed by Sudan Black B. In the cartilage matrix, a strong metachromasia was revealed by Toluidine Blue in the negatively-charged glycosaminoglycans. In the chondrocytes, glycogen granules were intensely positive to PAS method. Extracellular glycoproteins were also PAS positive in the basal membrane and in the region occupied by the lamina externa and reticular fibers surrounding each smooth muscle cells of the blood vessels. In the epithelial cells of the respiratory tract, acid and neutral mucins were histochemically detected by AB and PAS methods, respectively. Moreover, granules containing acid and neutral mucins were revealed in purple by AB and PAS concomitantly. In the sublingual gland sections, a distinct affinity of acid mucins by AB (in turquoise-blue) and collagen fibers by Picrosirius (in red) was obtained when these methods were combined. Although some routine dyes used in paraffin sections have showed a weak stain in historesin sections, our results showed that different dyes could be applied in GMA sections if modified staining procedures were assayed. Therefore, appropriate staining contrast and, thus, detection of one or different substances in a same section can be acquired in association to the good morphological resolution provided by GMA. PMID:14680922

Cerri, P S; Sasso-Cerri, E

2003-01-01

348

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

PubMed

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

Behr, Arno; Prez Gomes, Jessica

2011-01-01

349

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

PubMed Central

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

Ajjarapu, Srilatha; Shelef, Leora A.

1999-01-01

350

Potential of ethylenediaminedi(o-hydroxyphenylacetic acid) and N,N'-bis(hydroxybenzyl)ethylenediamine-N,N'-diacetic acid for the determination of metal ions by capillary electrophoresis.  

PubMed

Two aromatic polyaminocarboxylate ligands, ethylenediaminedi(o-hydroxyphenylacetic acid) (EDDHA) and N,N'-bis(hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED), were applied for the separation of transition and heavy metal ions by the ion-exchange variant of electrokinetic chromatography. EDDHA structure contains two chiral carbon centers. It makes it impossible to use the commercially available ligand. All the studied metal ions showed two peaks, which correspond to meso and rac forms of the ligand. The separation of metal-HBED chelates was performed using poly(diallyldimethylammonium) polycations in mixed acetate-hydroxide form. Simultaneous separation of nine single- and nine double-charged HBED chelates, including In(III), Ga(III), Co(II)-(III) and Mn(II)-(III) pairs demonstrated the efficiency of 40,000-400,000 theoretical plates. The separation of Co(III), Fe(III) complexes with different arrangements of donor groups and oxidation of Co(II), Mn(H), Fe(II) ions in reaction with HBED have been discussed. PMID:11009040

Krokhin, O V; Kuzina, O V; Hoshino, H; Shpigun, O A; Yotsuyanagi, T

2000-08-25

351

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

PubMed Central

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

Perez Gomes, Jessica

2011-01-01

352

Fabrication of nanoparticulate porous LaOF films through film growth and thermal decomposition of ion-modified lanthanum diacetate hydroxide.  

PubMed

This paper first reports fabrication of macro/nanotextured rare-earth oxyfluoride films. Usage of ion-modified lanthanum diacetate hydroxide (LDAH) as self-templates was successful in producing nanoparticulate lanthanum oxyfluoride (LaOF) films. LDAH template films were deposited on glass substrates through a chemical bath deposition in solutions composed of lanthanum acetate sesquihydrate, methanol, trifluoroacetic acid, and aqueous ammonia. The LDAH films had a unique, nestlike morphology owing to a two-dimensional hexagonal crystal growth. Modification of LDAH with trifluoroacetate ions led to formation of LaOF after pyrolyzing the template films at temperatures of 400-600 degrees C in air. The resultant LaOF films had a nanoparticulate porous microstructure, maintaining the morphology of the original LDAH template films. It was also successful to incorporate Eu3+ ions into LaOF through deposition of the LDAH film in a solution containing europium acetate tetrahydrate. The characteristic photoluminescence from Eu(3+) was observed with an ultraviolet-light excitation at 273 nm, indicating that Eu3+ was homogeneously distributed in LaOF host crystals. Thus the ion-modification of LDAH was also demonstrated to be a useful method for preparing nanostructured rare-earth oxyfluoride materials havingvarious cationic compositions. PMID:15875413

Hosono, Eiji; Fujihara, Shinobu; Kimura, Toshio

2004-04-27

353

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

SciTech Connect

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

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

2000-05-01

354

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

Microsoft Academic Search

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

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

2005-01-01

355

Comparison of ultra-widefield fluorescein angiography with the Heidelberg Spectralis noncontact ultra-widefield module versus the Optos Optomap  

PubMed Central

Purpose To compare ultra-widefield fluorescein angiography imaging using the Optos Optomap and the Heidelberg Spectralis noncontact ultra-widefield module. Methods Five patients (ten eyes) underwent ultra-widefield fluorescein angiography using the Optos panoramic P200Tx imaging system and the noncontact ultra-widefield module in the Heidelberg Spectralis HRA+OCT system. The images were obtained as a single, nonsteered shot centered on the macula. The area of imaged retina was outlined and quantified using Adobe Photoshop C5 software. The total area and area within each of four visualized quadrants was calculated and compared between the two imaging modalities. Three masked reviewers also evaluated each quadrant per eye (40 total quadrants) to determine which modality imaged the retinal vasculature most peripherally. Results Optos imaging captured a total retinal area averaging 151,362 pixels, ranging from 116,998 to 205,833 pixels, while the area captured using the Heidelberg Spectralis was 101,786 pixels, ranging from 73,424 to 116,319 (P = 0.0002). The average area per individual quadrant imaged by Optos versus the Heidelberg Spectralis superiorly was 32,373 vs 32,789 pixels, respectively (P = 0.91), inferiorly was 24,665 vs 26,117 pixels, respectively (P = 0.71), temporally was 47,948 vs 20,645 pixels, respectively (P = 0.0001), and nasally was 46,374 vs 22,234 pixels, respectively (P = 0.0001). The Heidelberg Spectralis was able to image the superior and inferior retinal vasculature to a more distal point than was the Optos, in nine of ten eyes (18 of 20 quadrants). The Optos was able to image the nasal and temporal retinal vasculature to a more distal point than was the Heidelberg Spectralis, in ten of ten eyes (20 of 20 quadrants). Conclusion The ultra-widefield fluorescein angiography obtained with the Optos and Heidelberg Spectralis ultra-widefield imaging systems are both excellent modalities that provide views of the peripheral retina. On a single nonsteered image, the Optos Optomap covered a significantly larger total retinal surface area, with greater image variability, than did the Heidelberg Spectralis ultra-widefield module. The Optos captured an appreciably wider view of the retina temporally and nasally, albeit with peripheral distortion, while the ultra-widefield Heidelberg Spectralis module was able to image the superior and inferior retinal vasculature more peripherally. The clinical significance of these findings as well as the area imaged on steered montaged images remains to be determined.

Witmer, Matthew T; Parlitsis, George; Patel, Sarju; Kiss, Szilard

2013-01-01

356

Staining of Internal Limiting Membrane in Macular Hole Surgery  

Microsoft Academic Search

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

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

2000-01-01

357

Analysis of Oxiclean: An Interesting Comparison of Percarbonate Stain Removers  

ERIC Educational Resources Information Center

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

Bracken, Jeffrey D.; Tietz, David

2005-01-01

358

ANEUPLOIDY TEST DEVELOPMENT: KINETOCHORE STAINING IN MAMMALIAN SYSTEMS  

EPA Science Inventory

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

359

Silver staining of fibrous neuroglia by means of physical development  

Microsoft Academic Search

A simple and reproducible technique for demonstrating fibrous neuroglia has been worked out on the basis of a new silver staining principle. It may be applied to formol-fixed material even after several years. The principle of it is as follows: In an acidic solution uranyl ions combine with or are adsorbed on fibrous neuroglia. The bound uranyl ions are replaced

F. Gallyas

1970-01-01

360

Nonfluorescent negative stain for alanopine dehydrogenase activity on starch gels.  

PubMed

A two-step procedure for negative staining NADH-dependent alanopine dehydrogenase (ALPDH) activity on starch gels is described, using nitroblue tetrasolium and phenasine methosulfate. This method gives a blue background with unreacted NADH, and white areas where ALPDH is located. Using this technique the first reliable evidence of the existence of true ALPDH isoenzymes is obtained. PMID:2409834

Manchenko, G P

1985-03-01

361

Incidence of homogeneously staining regions in non-Hodgkin lymphomas  

Microsoft Academic Search

We report a series of 86 non-Hodgkin's lymphomas (NHL) studied cytogenetically in which two cases with a homogeneously staining region (HSR) located on chromosome 19 and on chromosome 1, respectively, were observed. The low incidence detected (2.3%) suggests that HSR are rare events in NHL. An oncogenetic amplification study was performed with probes for genes that are currently known to

E. Arranz; M. Robledo; B. Martnez; J. Gallego; A. Romn; C. Rivas; J. Bentez

1996-01-01

362

Silver enhancement of polymerised diaminobenzidine: increased sensitivity for immunoperoxidase staining.  

PubMed Central

Unambiguous identification of lymphocytes is sometimes difficult because of weak immunostaining of the cell membrane immunoglobulins. A simple method of intensifying the diaminobenzidine (DAB) peroxidase reaction was therefore devised. Paraffin wax sections of formalin fixed tonsils and lymphomas were digested with trypsin and immunostained for kappa and lambda light immunoglobulin chains and CD3 antigen by various peroxidase linked detection systems. After reaction with hydrogen peroxide and DAB the sections were immersed in methenamine silver solution at 60 degrees C for three to seven minutes. The light brown stain on the cell membranes of the mantle zone lymphocytes became dark brown and the stronger stain of the plasma cells became black. Mantle zone B lymphocytes and CD3 positive T lymphocytes were precisely outlined even at low magnification and the lymphomas were easily classified as monoclonal or polyclonal. At high magnification, staining was clearer than with the immunogold-silver stain. Cryostat and paraffin wax sections of other tissues immunostained for various antigens showed similar intensification. Silver methenamine provides an easy means of increasing the sensitivity and visual impact of an immunoperoxidase/DAB reaction in any preparation. Images

Peacock, C S; Thompson, I W; Van Noorden, S

1991-01-01

363

PREFERENTIAL STAINING OF NUCLEIC ACIDCONTAINING STRUCTURES FOR ELECTRON MICROSCOPY  

Microsoft Academic Search

Oriented fibres of extracted nuclcohistone 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 tctroxide solution at pH 6, con- taining l0 -2 M Ca ++, and embedding in Araldite enabled sections of the fibres to be cut in which the

H. E. Huxley; G. ZUBAY

1961-01-01

364

Image analysis of dye stained patterns in soils  

NASA Astrophysics Data System (ADS)

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

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

2013-04-01

365

Fluorescence Changes during Conduction in Nerves Stained with Acridine Orange  

Microsoft Academic Search

Nerves from spider crabs and squid fluoresce when stained with Acridine Orange. The intensity of fluorescence increases during nerve conduction. Prolongation of the electric response in the squid axon is associated with a fluorescence change of similar duration. These findings suggest that the physicochemical properties of the macromolecules around the dye molecules in the nerve membrane drastically change during the

I. Tasaki; L. Carnay; R. Sandlin; A. Watanabe

1969-01-01

366

MODELING OF ALKANE EMISSIONS FROM A WOOD STAIN  

EPA Science Inventory

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

367

MODELING OF ALKANE EMISSIONS FROM A WOOD STAIN  

EPA Science Inventory

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

368

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

NASA Astrophysics Data System (ADS)

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.

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

2012-06-01

369

In-vivo buccal delivery of fluorescein isothiocyanate-dextran 4400 with glycodeoxycholate as an absorption enhancer in pigs.  

PubMed

Buccal delivery of fluorescein isothiocyanate labeled dextran 4400 (FD4) was investigated in-vivo in pigs. The delivery device consisted of an application chamber with a solution of FD4 and was adhered to the buccal mucosa for 4 h using an adhesive patch. A randomized crossover study including intravenous administration and buccal delivery without and with 10 mM sodium glycodeoxycholate (GDC) as absorption enhancer was performed in five pigs. After buccal administration, steady state plasma levels were rapidly reached. Coadministration of 10 mM GDC increased the absolute bioavailability of FD4 from 1.8 +/- 0.5% to 12.7 +/- 2.0%. Since FD4 is a macromolecular and hydrophilic compound such as peptide and protein drugs, buccal delivery would provide an adequate alternative to the parenteral administration of these drugs. PMID:8742934

Hoogstraate, A J; Verhoef, J C; Tuk, B; Pijpers, A; van Leengoed, L A; Verheijden, J H; Junginger, H E; Bodd, H E

1996-05-01

370

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.

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

2012-01-01

371

Photochemical initiation of thrombosis. Fluorescein angiographic, histologic, and ultrastructural alterations in the choroid, retinal pigment epithelium, and retina.  

PubMed

A new method of producing vascular occlusion, photochemical activation of intravenously injected rose bengal, was used to produce experimental thrombosis of the preretinal and choroidal blood vessels in rabbit eyes. Fluorescein angiography and light and electron microscopy were used to describe the resultant pathologic alterations over time. As early as one hour after treatment, the endothelium of both preretinal and choroidal blood vessels was severely damaged or completely obliterated, and platelet aggregates occluded the vascular lumina. Occlusion of the preretinal and choroidal blood vessels persisted for up to three days; however, endothelial regeneration and reperfusion had occurred in both vascular beds by seven days. In addition, the retinal pigment epithelium and myelin wings suffered ischemic damage. The retinal pigment epithelium began to recover by seven days, but the myelin wings appeared to be irreversibly damaged. PMID:3190547

Royster, A J; Nanda, S K; Hatchell, D L; Tiedeman, J S; Dutton, J J; Hatchell, M C

1988-11-01

372

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

PubMed

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

2012-06-01

373

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

374

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

SciTech Connect

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

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

1985-05-01

375

A staining procedure for micronucleus test using new methylene blue and acridine orange: specimens that are supravitally stained with possible long-term storage  

Microsoft Academic Search

The micronucleus test has been widely used as an in vivo cytogenetic test. It employs two different kinds of supravital staining methods which use either new methylene blue (N) and Giemsa (G) or acridine orange (AO). We have developed a new staining procedure for the preparation of specimens supravitally stained with possible long-term storage, using both N and AO. This

Tadakazu Sugihara; Shigeki Sawada; Atsushi Hakura; Yuji Hori; Kanako Uchida; Fumio Sagami

2000-01-01

376

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

377

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

378

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

379

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

380

A fluorescent Gram stain for flow cytometry and epifluorescence microscopy.  

PubMed

The fluorescent nucleic acid binding dyes hexidium iodide (HI) and SYTO 13 were used in combination as a Gram stain for unfixed organisms in suspension. HI penetrated gram-positive but not gram-negative organisms, whereas SYTO 13 penetrated both. When the dyes were used together, gram-negative organisms were rendered green fluorescent by SYTO 13; conversely, gram-positive organisms were rendered red-orange fluorescent by HI, which simultaneously quenched SYTO 13 green fluorescence. The technique correctly predicted the Gram status of 45 strains of clinically relevant organisms, including several known to be gram variable. In addition, representative strains of gram-positive anaerobic organisms, normally decolorized during the traditional Gram stain procedure, were classified correctly by this method. PMID:9647848

Mason, D J; Shanmuganathan, S; Mortimer, F C; Gant, V A

1998-07-01

381

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.

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

2012-01-01

382

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.

383

Complex III staining in blue native polyacrylamide gels.  

PubMed

For more than a decade now blue native polyacrylamide gel electrophoresis (BN-PAGE) has been used for the study of the oxidative phosphorylation (OXPHOS) complexes. Catalytic activities of complexes I, II, IV and V can be assessed, after separation by gel electrophoresis, by incubation of the BN-PAGE gel in specific staining solutions. However, until now, a reliable staining method for testing ubiquinol cytochrome c oxidoreductase (complex III) activity by BN-PAGE gel techniques was not available. In addition, spectrophotometric methods currently in use for detection of complex III deficiency in patients are not very sensitive. Here, we describe a newly developed diagnostic method for visualization of complex III activity by direct in-gel evaluation of ubiquinol cytochrome oxidoreductase activity. We validated the method by reporting the results in six patients with previously characterised complex III defects. PMID:21484424

Smet, Jol; De Paepe, Boel; Seneca, Sara; Lissens, Willy; Kotarsky, Heike; De Meirleir, Linda; Fellman, Vineta; Van Coster, Rudy

2011-06-01

384

The basis of differential staining of ascospores and vegetative cells of yeasts  

Microsoft Academic Search

An investigation of the staining reactions of ascospores from 10 species of yeasts with 19 different dyes, showed that differentiation of spores from vegetative cells by staining depends on their relative permeability properties. Three possible staining mechanisms are discussed and examined for consistency with the data. It was also discovered that ascospores of Hansenula saturnus, which are not stained by

M. S. Kelly; J. L. Gay

1969-01-01

385

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

Microsoft Academic Search

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

FENG CHEN; JING-RANG LU; BRIAN J. BINDER; YING-CHUN LIU; ROBERT E. HODSON

2001-01-01

386

Evacuation Traces Mini Challenge award: Innovative trace visualization staining for information discovery  

Microsoft Academic Search

Staining is a technique for categorizing time-varying spatial data; that is, data of things moving through space over time. In Staining, a stain is applied in either time or space, and the objects which move through the stain become marked. This technique and a research prototype demonstrating the technique were developed in response to the VAST 2008 Contest Mini-challenge: Evacuation

Dennis J. Bouvier; Britian Oates

2008-01-01

387

Impaired blood flow and arterio-venous shunting in human diabetic neuropathy: a novel technique of nerve photography and fluorescein angiography  

Microsoft Academic Search

SummaryNew techniques of sural nerve photography and fluorescein angiography which are able to provide an index of nerve blood flow have been developed. Under local anaesthetic, 3 cm of sural nerve was exposed at the ankle using an operating microscope. Without disturbing the epineurium, vessels were identified and photographed at a standard magnification ( 30). These were independently graded by

S. Tesfaye; N. Harris; J. J. Jakubowski; C. Mody; R. M. Wilson; I. G. Rennie; J. D. Ward

1993-01-01

388

The exogenous fluorophore, fluorescein, enables in vivo assessment of the gastrointestinal mucosa via confocal endomicroscopy: optimization of intravenous dosing in the dog model.  

PubMed

This study described the pharmacokinetics of the intravenous fluorophore, fluorescein, and aimed to evaluate its utility for use in upper gastrointestinal confocal endomicroscopy (CEM). Six healthy, mature, mixed-breed dogs were anesthetized and then dosed intravenously with fluorescein at 15 mg/kg. Blood samples were collected at predetermined time-points. Dogs were examined by upper gastrointestinal confocal endomicroscopy and monitored for adverse effects. Plasma fluorescein concentrations were measured using high-performance liquid chromatography (HPLC) with UV/Vis detection. Mean plasma concentration at 5 min was 57.6 18.2 mg/L, and plasma concentrations decreased bi-exponentially thereafter with a mean concentration of 2.5 mg/L 1.26 at 120 min. Mean terminal plasma elimination half-life (t? ) was 34.8 8.94 min, and clearance was 9.1 3.0 mL/kg/min. Apparent volume of distribution at steady-state was 0.3 0.06 L/kg. Fluorescein provided optimal fluorescent contrast to enable in vivo histologically equivalent evaluation of topologic mucosal morphology within the first 30 min following intravenous administration. Adverse effects were not observed. Based upon the calculated clearance, a constant rate infusion at a rate of 0.18 mg/kg/min is predicted to be adequate, following an initial loading dose (2 mg/kg), to maintain plasma concentration at 20 mg/L for optimal CEM imaging during the study period. PMID:23240692

Sharman, M J; Mansfield, C S; Whittem, T

2013-10-01

389

Fluorescence Quantum Yields and Their Relation to Lifetimes of Rhodamine 6G and Fluorescein in Nine Solvents: Improved Absolute Standards for Quantum Yields  

Microsoft Academic Search

Absolute fluorescence quantum yields are reported for the rhodamine 6G cation and the fluorescein dianion dyes in nine solvents. This information is combined with pre- viously reported fluorescence lifetimes to deduce radia- tive and nonradiative decay rates. Along the alcohol se- ries from methanol to octanol, rhodamine 6G displays an increasing radiative rate, in parallel with the square of the

Douglas Magde; Roger Wong; Paul G. Seybold

2002-01-01

390

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 (). The accumulation of fluorescence in the Caco-2 cell nucleus was observed mainly after the plateau phase of cell growth, indicating that permeated the nuclear envelope without nuclear-localizing tags. PMID:25000245

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

2014-07-22

391

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

392

Method and apparatus for staining immobilized nucleic acids  

SciTech Connect

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

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

2000-05-02

393

Lead-haematoxylin as a stain for endocrine cells  

Microsoft Academic Search

A modification of MacConaill's lead-haematoxylin has been found to stain several endocrine cells producing polypeptides and monoamines, particularly A and D cells of the pancreatic islet, thyroid C cells, gastro-intestinal enterochromaffin cells, gastric G and X cells, pituitary ACTH and MSH cells, adrenal medullary cells, and chemoreceptive cells of the carotid body. A careful comparison of the results of this

E. Solcia; C. Capella; G. Vassallo

1969-01-01

394

Y chromosome visibility in quinacrine-stained human spermatozoa  

Microsoft Academic Search

THE fluorescent dye, quinacrine, binds strongly to the Y chromosome and only to a lesser extent to the other chromosomes in human metaphase cell preparations1. A bright fluorescent spot (F body), which is presumed to be the Y chromosome, is clearly visible in stained interphase cells from various tissues from the human male2-5 and in mature spermatozoa6,7. Spermatozoa bearing one

A. M. Roberts; H. Goodall

1976-01-01

395

Method and apparatus for staining immobilized nucleic acids  

DOEpatents

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

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

2000-01-01

396

Use of commercially available monoclonal antibodies for immunoenzyme double staining  

Microsoft Academic Search

SummaryAn immunoenzyme double-staining method for the simultaneous detection of two cellular epitopes, using commercially available mouse monoclonal antibodies, is described. The method employs a combination of the suppression of endogenous biotin and two successive indirect techniques with a blocking step in between. The first indirect method involves an unlabelled monoclonal antibody followed by an alkaline phosphatase-conjugated goat anti-mouse immunoglobulin. After

C. M. Van Der Loos; J. J. Van Den Oord; P. K. Das; H. J. Houthoff

1988-01-01

397

Improved avidinbiotinperoxidase complex (ABC) staining  

Microsoft Academic Search

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

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

1988-01-01

398

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

399

Routine papillomavirus antigen staining of cervical punch biopsy specimens.  

PubMed Central

Immunocytochemical staining for papillomavirus antigen was carried out on 1147 consecutive cervical punch biopsy specimens over 12 months. Of 876 cases with cervical intraepithelial neoplasia (CIN) 351, were antigen positive and of 49 cases with histological evidence of human papillomavirus (HPV) infection but no CIN, 14 were positive. There were 204 cases reported to be normal on routine histological examination and 12 cases reported to show features suggestive but not diagnostic of HPV infection. Of the normal group, 24 (12%) were antigen positive and of the equivocal group, two were positive. In 122 of the normal or equivocal groups cytological examination was repeated at the time of colposcopy, and dyskaryosis was reported in 36. In only four cases was disease shown by HPV antigen staining when there was no diagnostic histological or cytological abnormality. HPV antigen staining assists in the recognition of the range of histological changes associated with productive HPV infection but is an insensitive test and has only limited value in supplementing histological and cytological examinations as a diagnostic aid in routine colposcopic pathology. Images Fig 1 Fig 1 Fig 2 Fig 2

Jenkins, D; Tay, S K; Maddox, P H

1987-01-01

400

``Gold corrosion'': red stains on a gold Austrian Ducat  

NASA Astrophysics Data System (ADS)

Stains of different colours have been observed on historic and modern gold coins in several countries. An Austrian Ducat at the Kunsthistorisches Museum in Vienna has developed some red spots on its surface over the years. The same defects have also been observed in modern coins of higher gold purity. The spots have been examined by OM, SEM, EDS, XPS and AES. Optical microscopy showed that ``red'' defects exhibit in fact a nuance of colours. The surface analysis put in evidence the presence in the stains, in addition to gold, of silver and sulphur. The values of the modified Auger parameter ?' of silver correspond to those of Ag2S; thus, it can be assumed that the stains are composed of silver sulphide (Ag2S). It was not possible to determine whether the presence of silver on the surface is due to segregation towards the surface or to external particles of silver embedded in the matrix. Depth profiling performed on modern coins suffering from the same problem allowed us to demonstrate that the nuance of colours is due to the inhomogeneous thickness of the spots. Moreover, it was demonstrated that spots are formed by two layers: an outer layer of silver sulphide and an inner layer of silver.

Gusmano, G.; Montanari, R.; Kaciulis, S.; Montesperelli, G.; Denk, R.

401

Congenital ichthyosiform erythroderma: particulate staining pattern of TGK.  

PubMed

A case of late onset non-bullous congenital ichthyosiform erythroderma (CIE) was studied. This patient was not born as a collodion baby and did not have skin abnormalities until 9-10 years of age. She gradually developed erythroderma and fine scales, callosities of her feet, and a mild ectropion. Since recent work has revealed that in the majority of CIE patients, transglutaminase (TGK) is distributed in the cytoplasm of granular cells and horny cells (11), TGK was studied in our case. It was found that TGK was distributed along the cell periphery of horny cells and also in the cytoplasm of granular cells. In the control skins, TGK was stained along the cell periphery of horny cells and granular cells. The marginal band formation was normal. Involucrine and loricrin, the building materials of the marginal band whose-cross-linking is mediated by TGK, were normally stained in the upper epidermis. Cytoplasmic TGK of granular cells and normal development of the marginal band may serve as a helpful diagnostic marker of CIE, particularly because the often confusing collodion baby of lamellar ichthyosis may lack TGK staining and the marginal band altogether. PMID:10659499

Hashimoto, K; Tanaka, K; Misra, I; Shwayder, T; Moiin, A

1999-12-01

402

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

403

[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

404

Identification of homogeneously staining regions in leukemia patients  

PubMed Central

Homogeneously staining regions (HSR) or double minute chromosomes (dmin) are autonomously replicating extra-chromosomal elements that are frequently associated with gene amplification in a variety of cancers. The diagnosis of leukemia patients was based on characterization of the leukemic cells obtained from bone marrow cytogenetics. This study report two cases, one with Acute Myeloblastic Leukemia without maturation (AML-M1), aged 23-year-old female, and the other with chronic myelogenous leukemia (CML)-blast crisis, a 28-year-old female associated with double minute chromosomes. Most cases of acute myeloid leukemia with dmin in the literature (including our cases) have been diagnosed as having acute myeloid leukemia.

Moghadam, Mohammad Heydarian; Movafagh, Abolfazl; Omrani, MirDavood; Ghanati, Kiandokht; Hashemi, Mehrdad; Poursafavi, Farhikhteh; Darvish, Hossein; Abdolahi, Davood Zare; Gholami, Milad; Heidari Rostamy, Mohammad Reza; Safari, Shamsi; HaghNejad, Leyla; Darehgazani, Reyhaneh; Naeini, Niloofar Safavi; Motlagh, Mehdi Ghandehari; Amani, Davar

2013-01-01

405

[Accidental staining of corneal nerves by methylene blue].  

PubMed

A 10-year-old child presented after accidental exposure of the left eye to a blue hair dye containing methylene blue. Mild ocular surface changes and a selective blue staining of the usually invisible corneal nerve fibre bundles were present. Corneal sensitivity was reduced. Despite copious lubrication a transient neurotrophic keratitis developed which did not resolve until corneal sensitivity became normal 2 weeks later. Association of mild chemical burns with neurotrophic keratitis is unusual but is of high clinical relevance as keratitis is a vision-threatening complication. PMID:23288315

Peter, S; Reichart, E; Poyntner, L; Mennel, S

2013-09-01

406

Automated Analysis of PIN-4 Stained Prostate Needle Biopsies  

NASA Astrophysics Data System (ADS)

Prostate Needle biopsies are stained with the PIN-4 marker cocktail to help the pathologist distinguish between HGPIN and adenocarcinoma. The correct interpretation of multiple IHC markers can be challenging. Therefore we propose the use of computer aided diagnosis algorithms for the identification and classification of glands in a whole slide image of prostate needle biopsy. The paper presents the different issues related to the automated analysis of prostate needle biopsies and the approach taken by BioImagene in its first generation algorithms.

Sabata, Bikash; Babenko, Boris; Monroe, Robert; Srinivas, Chukka

407

Staining Characteristics and Antiviral Activity of Sulforhodamine B and Lissamine Green B  

Microsoft Academic Search

Purpose. Fluorescein and rose bengal are dyes used routinely in the examination of the ocular surface. As part of an ongoing search for a superior ophthalmic dye with optimal specificity and sensitivity and a lack of interference with subsequent viral cultures, and as part of studies that use chemical dyes to understand better the pathophysiology of ocular surface disorders, the

James Chodosh; Richard D. Dix; R. Clark Howell; William G. Stroop; Scheffer C. G. Tseng

408

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

PubMed

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

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

2013-01-01

409

Multiple binding sites of fluorescein isothiocyanate moieties on myoglobin: photophysical heterogeneity as revealed by ground- and excited-state spectroscopy.  

PubMed

Fluorescein isothiocyanate (FITC)-myoglobin conjugates were synthesized with a binding stoichiometry of one to three fluorophores per protein. FITC binding sites were determined by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS). Five lysine residues and the N-terminal amino group were identified as preferential binding sites. The ground and excited-state absorption spectra and the fluorescence decay of the conjugates in the native and denatured state of the carrier protein were analyzed. For comparison, unbound FITC and FITC covalently bound to a polysaccharide (dextran) were studied. For FITC, FITC-dextran and the FITC-myoglobin conjugates, only one FITC absorption peak was obtained in the ground state spectrum. Similarly, the excited state absorption (ESA) spectra of unbound FITC and of FITC-dextran showed only one single maximum whereas two maxima were detected for the native FITC-myoglobin conjugates. One of these sub-bands disappeared following urea treatment of the conjugate. We conclude that ESA measurements of extrinsic fluorophores on proteins can be used to monitor different micro-environments of the fluorophore and to distinguish between different conformational states of the labeled protein. This method can be a useful tool for analysing coexisting protein conformations. PMID:12167317

Grunwaldt, Gisela; Haebel, Sophie; Spitz, Christian; Steup, Martin; Menzel, Ralf

2002-07-01

410

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

PubMed

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

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

2015-01-01

411

Evaluation of a Novel, Non Contact, Automated Focal Laser with Integrated (NAVILAS) Fluorescein Angiography for Diabetic Macular Edema  

PubMed Central

Purpose: To evaluate the procedural experience and complications of a novel integrated laser delivery system (NAVILAS; OD-OS Teltow, Germany) that combines automated laser delivery with color fundus photography, fluorescein angiography (FA), fundus autofluorescence (FAF) and infrared imaging with a frequency doubled YAG laser. Materials and Methods: This prospective study evaluated surgical experience with the NAVILAS automated photocoagulation system for the treatment of patients with diabetic macular edema (DME). Subjective assessment of the accuracy of laser spot placement and postoperative complications were documented. Results: Twelve patients (7 males, 5 females) were enrolled in this pilot study. Five patients were phakic and 7 were pseudophakic. Image overlays and the tracking system allowed accurate delivery of laser spots of varying size, duration and power. None of the patients reported any pain and tolerated the procedure well. No complications were reported in the study. Conclusion: In this pilot study, the NAVILAS system allowed accurate laser spot placement with no complications in patients with DME. However a larger sample with longer follow up is required to determine the safety of this procedure.

Chalam, Kakarla V.; Murthy, Ravi K.; Brar, Vikram; Radhakrishnan, Ravi; Khetpal, Vijay; Grover, Sandeep

2012-01-01

412

Fluorescence turn-on detection of iodide, iodate and total iodine using fluorescein-5-isothiocyanate-modified gold nanoparticles.  

PubMed

Selective turn-on fluorescence detection of I(-) was accomplished using fluorescein isothiocyanate-decorated gold nanoparticles (FITC-AuNPs). FITC molecules, which fluoresce strongly in an alkaline solution, were severely quenched when they were attached to the surface of AuNPs through their isothiocyanate group. Upon the addition of I(-), FITC molecules were detached because of I(-) adsorption on the surface of AuNPs. As a result, released FITC molecules were restored to their original fluorescence intensity. Because I(-) has a higher binding affinity to the surface of Au than do Br(-), Cl(-), or F(-), the FITC-AuNPs obviously have a higher selectivity toward I(-) than toward these other anions. Meanwhile, after IO(3)(-) was reduced to I(-) with ascorbic acid, the detection of IO(3)(-) was successfully achieved using the FITC-AuNPs. Under an optimum pH and AuNP concentration, the lowest detectable concentrations of I(-) and IO(3)(-) using this probe were 10.0 and 50.0 nM, respectively. The FITC-AuNPs provide a number of advantages, including easy preparation, selectivity, sensitivity, and low cost. This unique probe was applied to an analysis of the total iodine in edible salt and seawater. PMID:19768221

Chen, Yi-Ming; Cheng, Tian-Lu; Tseng, Wei-Lung

2009-10-01

413

Basal values, intra-day and inter-day variations in tear film osmolarity and tear fluorescein clearance.  

PubMed

Abstract Purpose: The aim of this study was to determine the normal inter-day and intra-day variations in tear film osmolarity and the tear fluorescein clearance test (T-FCT) in healthy subjects. Methods: Tear samples from 24 young, healthy adults were collected from 11:00?AM to 1:00?PM (midday) and 5:00?PM to 7:00?PM (evening) on three non-consecutive days. Tear osmolarity measurement and the T-FCT were performed to assess the basal values and inter-day and intra-day variations of the test results. A freezing point depression osmometer was used to analyze the tear osmolarity, and the T-FCT was performed using a fluorophotometer. Results: The mean osmolarity value was 270??4.4?mOsm/l and the mean T-FCT result was 2.97??0.17 fluorescence arbitrary units. The inter-day or intra-day tear osmolarity values did not differ significantly. The T-FCT results varied significantly during the day, with significantly (p?=?0.0004) higher results in the evening; no significant differences were found in the inter-day analysis. Conclusions: Tear osmolarity was unaffected by intra-day variations; however, the T-FCT showed an inter-day variation, which indicated that the time of day when the test is performed must be considered when it is used to evaluate the diagnosis of dry eye disease, disease progression or therapeutic effectiveness. PMID:24400688

Garca, Noelia; Tesn, Marisa; Enrquez-de-Salamanca, Amalia; Mena, Laura; Sacristn, Amelia; Fernndez, Itziar; Calonge, Margarita; Gonzlez-Garca, Mara J

2014-07-01

414

The ultimate Wright-Giemsa stain: 60 years in the making.  

PubMed

Abstract Wright-Giemsa staining is a common procedure that is performed routinely in hematology laboratories. Consistency in intra-laboratory staining quality is essential for accurate morphological interpretation of blood smears. Although the Wright-Giemsa stain can be challenging to perform, the methods illustrated here have provided consistent, high quality stains in the Special Hematology Laboratory at the University of Minnesota for over half a century. We outline methods for collecting blood specimens, preparing the slides and performing a Wright-Giemsa stain using our combination of reagents. Various techniques that have been passed down in our laboratory for troubleshooting suboptimally stained specimens are shared as well. PMID:21395491

Dunning, K; Safo, A O

2011-04-01

415

Antibody staining in C. elegans using "freeze-cracking".  

PubMed

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

Duerr, Janet S

2013-01-01

416

Determination of (4E,6E,12E)-Tetradecatriene-8,10-diyne-1,3-diyl Diacetate in Rat Plasma and Tissues by HPLC-UV Method and Their Application to a Pharmacokinetic and Tissue Distribution Study  

PubMed Central

In China Atractylodis Rhizoma is widely used for the treatment of rheumatic diseases and digestive disorders. Stir-frying with wheat bran is the most common processing method. In order to clarify the influence of processing on pharmacological properties of Atractylodis Rhizoma, an investigation was carried out to compare the pharmacokinetics and tissue distribution of typical constituent after oral administration of raw Atractylodis Rhizoma and processed ones. A simple, rapid, and sensitive high performance liquid chromatography with UV detection was developed and validated for the determination of (4E,6E,12E)-tetradecatriene-8,10-diyne-1,3-diyl diacetate in rat plasma. A chromatography was carried out on Diamonsil C18 (250 4.6?mm; 5??m) analytical column, using a mobile phase which consisted of acetonitrile and 0.1% phosphoric acid water (60?:?40, v/v) at a flow rate of 1.0?mLmin?1. The wavelength was set at 336?nm. The LLOQ of (4E,6E,12E)-tetradecatriene-8,10-diyne-1,3-diyl diacetate was 0.00143??gmL?1. Both accuracy and precision were satisfactory. The pharmacokinetic results showed that the Tmax? was 1 hour in advance and the Cmax? was increased after processing. Tissue distribution showed that the highest level was in spleen. And the concentrations in the spleen were increased after stir-frying with bran.

Huo, Yan; Liu, Yu-Qiang; Bai, Zhong-Xu; Cai, Qian

2014-01-01

417

Effect of antihypertensive treatment on blood-retinal barrier permeability to fluorescein in hypertensive Type 1 (insulin-dependent) diabetic patients with background retinopathy  

Microsoft Academic Search

SummaryThe effect of antihypertensive treatment on blood-retinal barrier leakage of fluorescein in background retinopathy was studied in nine hypertensive Type 1 (insulin-dependent) diabetic patients suffering from nephropathy. The patients were investigated before and after 7 (3 to 13) months of treatment with captopril (n=8; 25 to 100 mg daily) and a diuretic, either frusemide (n=4; 80 to 200 mg daily)

H.-H. Parving; M. Larsen; E. Hommel; H. Lund-Andersen

1989-01-01

418

Hot water postprocess pasteurization of cook-in-bag turkey breast treated with and without potassium lactate and sodium diacetate and acidified sodium chlorite for control of Listeria monocytogenes.  

PubMed

Surface pasteurization and food-grade chemicals were evaluated for the ability to control listeriae postprocess on cook-in-bag turkey breasts (CIBTB). Individual CIBTB were obtained directly from a commercial manufacturer and surface inoculated (20 ml) with a five-strain cocktail (ca. 7.0 log) of Listeria innocua. In each of two trials, the product was showered or submerged for up to 9 min with water heated to 190, 197, or 205 degrees F (ca. 87.8, 91.7, or 96.1 degrees C) in a commercial pasteurization tunnel. Surviving listeriae were recovered from CIBTB by rinsing and were then enumerated on modified Oxford agar plates following incubation at 37 degrees C for 48 h. As expected, higher water temperatures and longer residence times resulted in a greater reduction of L. innocua. A ca. 2.0-log reduction was achieved within 3 min at 205 and 197 degrees F and within 7 min at 190 degrees E In related experiments, the following treatments were evaluated for control of Listeria monocytogenes on CIBTB: (i) a potassium lactate-sodium diacetate solution (1.54% potassium lactate and 0.11% sodium diacetate) added to the formulation in the mixer and 150 ppm of acidified sodium chlorite applied to the surface with a pipette, or (ii) a potassium lactate-sodium diacetate solution only, or (iii) no potassium lactate-sodium diacetate solution and no acidified sodium chlorite. Each CIBTB was inoculated (20 ml) with ca. 5 log CFU of a five-strain mixture of L. monocytogenes and then vacuum sealed. In each of two trials, half of the CIBTB were exposed to 203 degrees F water for 3 min in a pasteurization tunnel, and the other half of the CIBTB were not; then, all CIBTB were stored at 4 degrees C for up to 60 days, and L. monocytogenes was enumerated by direct plating onto modified Oxford agar. Heating resulted in an initial reduction of ca. 2 log CFU of L. monocytogenes per CIBTB. For heated CIBTB, L. monocytogenes increased by ca. 2 log CFU per CIBTB in 28 (treatment 1), 28 (treatment 2), and 14 (treatment 3) days. Thereafter, pathogen levels reached ca. 7 log CFU per CIBTB in 45, 45, and 21 days for treatments 1, 2, and 3, respectively. In contrast, for nonheated CIBTB, L. monocytogenes levels increased from ca. 5 log CFU per CIBTB to ca. 7 log CFU per CIBTB in 28, 21, and 14 days for treatments 1, 2, and 3, respectively. Lastly, in each of three trials, we tested the effect of hot water (203 degrees F for 3 min) postprocess pasteurization of inoculated CIBTB on the lethality of L. monocytogenes and validated that it resulted in a 1.8-log reduction in pathogen levels. Collectively, these data establish that hot water postprocess pasteurization alone is effective in reducing L. monocytogenes on the surface of CIBTB. However, as used in this study, the potassium lactate-sodium diacetate solution and acidified sodium chlorite were only somewhat effective at controlling the subsequent outgrowth of this pathogen during refrigerated storage. PMID:16416899

Luchansky, John B; Cocoma, George; Call, Jeffrey E

2006-01-01

419

Dual DNA Staining Assessment of Bovine Sperm Viability Using SYBR-14 and Propidium Iodide  

Microsoft Academic Search

A new membrane-permeant DNA stain, SYBR-1 4, was used in combination with propidium iodide (P1)to estimate the pro- portion of living sperm in bovine semen. The SYBR-14 stained living sperm while P1 only stained degenerate cells that had lost their membrane integrity. Staining with SYBR-1 4 resulted in the nuclei of living sperm fluorescing bright green. Aliquots containing nearly all

D. L. GARNER; L. A. JOHNSON; S. T. YUE; B. L. ROTH

420

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 714%. The proposed method is straightforward and can be applied to standardized microtiter plates.

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

2003-01-01

421

Immunocytochemical staining of cholinergic amacrine cells in rabbit retina.  

PubMed

Cholinergic neurons of rabbit retina were labelled with an antibody against choline acetyltransferase, the synthesizing enzyme for acetylcholine. Two populations of cells are immunoreactive. Type a cell bodies lie in the inner nuclear layer (INL), their dendrites branching narrowly in sublamina a of the inner plexiform layer (IPL), while type b cell bodies lie in the ganglion cell layer (GCL) with dendrites branching in sublamina b of the IPL. The irregular networks of clustered immunoreactive dendrites are similar, but not identical, in the two sublaminae. Type b cells are more numerous than type a cells in central retina. No axons were stained. It appears that the immunoreactive neurons are normally placed and displaced starburst/cholinergic amacrine cells. PMID:3300857

Famiglietti, E V; Tumosa, N

1987-06-16

422

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

423

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

2013-04-29

424

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

425

NF93-136 Chemical Spots, Stains and Discoloration of Home Furnishings  

Microsoft Academic Search

We live in a world of chemicals. Unfortunately, some of the characteristics that make household chemical products the most useful are the same qualities that lead to trouble when these products are carelessly handled. A chemical stain or spot is a serious kind of stain that is appearing with increasing frequency and is different from ordinary stains. This type of

Shirley Niemeyer

1993-01-01

426

Fuchsin fluorescence in Mycobacterium tuberculosis: The ZiehlNeelsen stain in a new light  

Microsoft Academic Search

Tuberculosis (TB) is often diagnosed by observation of reddish pink fuchsin-stained Mycobacterium tuberculosis (MTB) in ZiehlNeelsen (ZN) stained smears by transmitted light microscopy. MTB too faintly stained with fuchsin to be seen by transmitted light may be detected by their green-excited orange-red fluorescence; this finding may be clinically relevant.

Howard M. Shapiro; Thomas Hnscheid

2008-01-01

427

Digital Staining of Unstained Pathological Tissue Samples through Spectral Transmittance Classification  

Microsoft Academic Search

Histological structures of a pathological tissue sample convey information relevant to the diagnosis of the disease that might have afficted the person. To reveal the morphology of these structures clearly, pathological tissues are stained. In this paper, a digital staining methodology for pathological tissue samples is introduced. Digital staining implies the application of digital processing techniques to transform the image

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

2005-01-01

428

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

429

Fluoresceination of FepA during Colicin B Killing: Effects of Temperature, Toxin and TonB  

PubMed Central

We studied the reactivity of 35 genetically engineered Cys sulfhydryl groups at different locations in Escherichia coli FepA. Modification of surface loop residues by fluorescein maleimide (FM) was strongly temperature-dependent in vivo, whereas reactivity at other sites was much less affected. Control reactions with bovine serum albumin showed that the temperature dependence of loop residue reactivity was unusually high, indicating that conformational changes in multiple loops (L2, L3, L4, L5, L7, L8, L10) transform the receptor to a more accessible form at 37 C. At 0 C colicin B binding impaired or blocked labeling at 8 of 10 surface loop sites, presumably by steric hindrance. Overall, colicin B adsorption decreased the reactivity of more than half of the 35 sites, in both the N - and C- domains of FepA. However, colicin B penetration into the cell at 37 C did not augment the chemical modification of any residues in FepA. The FM modification patterns were similarly unaffected by the tonB locus. FepA was expressed at lower levels in a tonB host strain, but when we accounted for this decrease its FM-labeling was comparable whether TonB was present or absent. Thus we did not detect TonB-dependent structural changes in FepA, either alone or when it interacted with colicin B at 37 C. The only changes in chemical modification were reductions from steric hindrance when the bacteriocin bound to the receptor protein. The absence of increases in the reactivity of N-domain residues argues against the idea (Devanathan and Postle, Mol. Microbiol. 65: 441453, 2007) that the colicin B polypeptide traverses the FepA channel.

Smallwood, Chuck R.; Marco, Amparo Gala; Xiao, Qiaobin; Trinh, Vy; Newton, Salete M. C.; Klebba, Phillip E.

2009-01-01

430

Peripheral retinal ischaemia, as evaluated by ultra-widefield fluorescein angiography, is associated with diabetic macular oedema  

PubMed Central

Purpose To determine the relationship between retinal ischaemia and the presence of macular oedema (DMO) in patients with diabetic retinopathy (DR) using ultra-widefield fluorescein angiography (UWFA) imaging. Methods A retrospective review of 122 eyes of 70 treatment-nave diabetic patients who underwent diagnostic UWFA using the Optos 200Tx imaging system. Two independent, masked graders quantified the area of retinal ischaemia. Based on clinical examination and optical coherence tomography (OCT), each patient was given a binary classification as either having DMO or no DMO. McNemar's test (with Yates' correction as indicated) and a two-sample test of proportions were used to determine the relationship between DMO and ischaemia for binary and proportional data, respectively. Linear and logistic models were constructed using generalised estimating equations to test relationships between independent variables, covariates and outcomes while controlling for inter-eye correlation, age, gender, haemoglobin A1c, mean arterial pressure and dependence on insulin. Results Seventy-six eyes (62%) exhibited areas of retinal ischaemia. There was a significant direct correlation between DMO and peripheral retinal ischaemia as seen on UWFA (p<0.001). In addition, patients with retinal ischaemia had 3.75 times increased odds of having DMO compared with those without retinal ischaemia (CI 1.26 to 11.13, p<0.02). Conclusion Retinal ischaemia is significantly correlated with DMO in treatment-nave patients with DR. UWFA is a useful tool for detecting peripheral retinal ischaemia, which may have direct implications in the diagnosis, follow-up and treatment such as targeted peripheral photocoagulation.

Wessel, Matthew M; Nair, Nandini; Aaker, Grant D; Ehrlich, Joshua R; D'Amico, Donald J

2012-01-01

431

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.

Ankle, Madhuri R; Joshi, Priya S

2011-01-01

432

Coloring-decoloring behavior of amphiphilic fluoroalkyl end-capped N-(1,1-dimethyl-3-oxobutyl)acrylamide--acryloylmorpholine cooligomer/fluorescein nanocomposites in protic and aprotic solvents.  

PubMed

Amphiphilic fluoroalkyl end-capped N-(1,1-dimethyl-3-oxobutyl)acrylamide-acryloylmorpholine cooligomer/fluorescein nanocomposites afforded brilliant yellow-colored solutions in not only protic solvents such as methanol and ethanol but also protic-like solvents such as dichloromethane and 1,2-dichloroethane, respectively. However, the corresponding non-fluorinated cooligomer/fluorescein composites and parent fluorescein gave the colorless solutions under similar conditions. On the other hand, unexpectedly, such brilliant yellow-colored solutions provided by these fluorinated nanocomposites completely disappeared in aprotic solvents such as N,N-dimethylformamide, dimethyl sulfoxide, and tetrahydrofuran. Thus, these fluorinated fluorescein nanocomposites can exhibit a coloring-decoloring behavior through solvatochromic response. PMID:22484165

Sawada, Hideo; Izumi, Shunsuke; Sasazawa, Kazuo; Yoshida, Masato

2012-07-01

433

A rapid method for staining inclusions of Chlamydia psittaci and Chlamydia trachomatis.  

PubMed

A new staining method was developed for the detection of inclusions of Chlamydia psittaci and Chlamydia trachomatis inclusions in cell cultures. Using a combination of methyl green and neutral red stains and washing at pH 5.0, inclusions were stained red while cell cytoplasm was pale pink and cell nuclei were pale green. The method was significantly better than Giemsa staining and comparable to immunofluorescence for detecting C psittaci inclusions. Its sensitivity for detection C trachomatis inclusions by dark field microscopy was similar to that of Giemsa staining. PMID:6177720

Woodland, R M; Malam, J; Darougar, S

1982-06-01

434

Contrast staining of Trichinella spiralis larvae in fresh, frozen and formalin preserved muscles.  

PubMed

Fresh, frozen-thawed and formalin-preserved muscle samples heavily infected with Trichinella larvae were cut into several pieces and stained with Giemsa and Leishman and the reference Haematoxylineosin (H & E) stain. Observation under microscope revealed that both muscle larvae and nurse cells in fresh and formalin preserved specimens appeared as purplish blue structures contrasting with the pinkish color of non-infected muscle fibers in both Giemsa and Leishman stains. These findings were confirmed in H & E stained samples. However, frozen samples did not show contrast stain. PMID:18383802

Ali, Nehad M; Thabet, Hala S

2007-12-01

435

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

PubMed Central

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

Cherkezyan, Lusik; Subramanian, Hariharan; Stoyneva, Valentina; Rogers, Jeremy D.; Yang, Seungmoo; Damania, Dhwanil; Taflove, Allen; Backman, Vadim

2012-01-01

436

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 0C. 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 ( hour to 2 hours, at 0C or 20C), 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.

Huxley, H. E.; Zubay, G.

1961-01-01

437

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

PubMed

Visualization of bacteria in living plant cells and tissues is often problematic due to lack of stains that pass through living plant cell membranes and selectively stain bacterial cells. In this article, we report the use of 3,3'-diaminobenzidine tetrachloride (DAB) to stain hydrogen peroxide associated with bacterial invasion of eukaryotic cells. Tissues were counterstained with aniline blue/lactophenol to stain protein in bacterial cells. Using this staining method to visualize intracellular bacterial (Burkholderia gladioli) colonization of seedling roots of switch grass (Panicum virgatum), we compared bacterial free seedling roots and those inoculated with the bacterium. To further assess application of the technique in multiple species of vascular plants, we examined vascular plants for seedling root colonization by naturally occurring seed-transmitted bacteria. Colonization by bacteria was only observed to occur within epidermal (including root hairs) and cortical cells of root tissues, suggesting that bacteria may not be penetrating deeply into root tissues. DAB/peroxidase with counter stain aniline blue/lactophenol was effective in penetration of root cells to selectively stain bacteria. Furthermore, this stain combination permitted the visualization of the bacterial lysis process. Before any evidence of H2 O2 staining, intracellular bacteria were seen to stain blue for protein content with aniline blue/lactophenol. After H2 O2 staining became evident, bacteria were often swollen, without internal staining by aniline blue/lactophenol; this suggests loss of protein content. This staining method was effective for seedling root tissues; however, it was not effective at staining bacteria in shoot tissues due to poor penetration. Microsc. Res. Tech. 77:566-573, 2014. 2014 Wiley Periodicals, Inc. PMID:24825573

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

2014-08-01

438

Diffuse reflectance FTIR of stains on grit blasted metals  

SciTech Connect

Diffuse reflectance mid-infrared Fourier transform (DRIFT) spectroscopy has been applied to the detection of oil contamination on grit-blasted metals. The object of this application is to detect and discriminate between silicone and hydrocarbon oil contamination at levels approaching 10 mg m{sup {minus}2}. A portable FTIR spectrometer with dedicated diffuse reflectance optics was developed for this purpose. Using translation devices positioned by instructions from the spectrometer operating system, images of macroscopic substrates were produced with millimeter spatial resolution. The pixels that comprise an image are each a full mid-infrared spectrum with excellent signal-to-noise, each determined as individual files and uniquely saved to disc. Reduced spectra amplitudes, based on peak height, area, or other chemometric techniques, mapped as a function of the spatial coordinates of the pixel are used to display the image. This paper demonstrates the application of the technique to the analysis of stains on grit-blasted metals, including the calibration of the method, the inspection of substrates, and the migration of oil contamination. {copyright} {ital 1998 American Institute of Physics.}

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

1998-06-01

439

Diffuse reflectance FTIR of stains on grit blasted metals  

SciTech Connect

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

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

1997-08-09

440

An automatic nonrigid registration for stained histological sections.  

PubMed

Automatic computer-based analyses of histological sections which are differently stained require that they are related to each other. Most registration methods are only able to perform rigid-body motion and are sensitive to noise and artifacts. Histological images, however, are accompanied by several artifacts and different contrasts, which require a nonrigid registration. In this paper, we present a hierarchical nonrigid registration algorithm able to align images, which contain minor image artifacts. The algorithm requires no a priori knowledge of the true image. The hierarchical design of the algorithm enhances robustness and accuracy, and saves computational costs. The proposed algorithm is decomposed into a fast, coarse, rigid registration step and a slower, but finer, nonrigid step. For the coarse registration, we use image pyramids, while for the second step, we combine a point-based registration with an elastic thin-plate spline interpolation. Accuracy tests, performed for 20 histological images obtained from human arteries, have shown that the error measure is acceptable, and that the image noise does not cause a problem. The associated convergence rate of the mean pixel displacement error during the rigid and nonrigid registrations is satisfying. The algorithm can be applied to various multicontrast elastic registration problems in medical imaging and may be extended to three dimensions. PMID:15825482

Auer, Martin; Regitnig, Peter; Holzapfel, Gerhard A

2005-04-01

441

Local Histograms for Classifying H&E Stained Tissues  

PubMed Central

We introduce a rigorous mathematical theory for the analysis of local histograms, and consider the appropriateness of their use in the automated classification of textures commonly encountered in images of H&E stained tissues. We first discuss some of the many image features that pathologists indicate they use when classifying tissues, focusing on simple, locally-defined features that essentially involve pixel counting: the number of cells in a region of given size, the size of the nuclei within these cells, and the distribution of color within both. We then introduce a probabilistic, occlusion-based model for textures that exhibit these features, in particular demonstrating how certain tissue-similar textures can be built up from simpler ones. After considering the basic notions and properties of local histogram transforms, we then formally demonstrate that such transforms are natural tools for analyzing the textures produced by our model. In particular, we discuss how local histogram transforms can be used to produce numerical features that, when fed into mainstream classification schemes, mimic the baser aspects of a pathologist's thought process.

Massar, M.L.; Bhagavatula, R.; Fickus, M.; Kovacevic, J.

2010-01-01

442

Ultrafast dynamics of epicocconone, a second generation fluorescent protein stain.  

PubMed

Femtosecond upconversion experiment has been carried out for epicocconone and its butylamine adduct in acetonitrile and tert-butanol. An ultrafast component is found to dominate the decay of fluorescence of epicocconone in acetonitrile solution. Upon reacting with butylamine, a model for the epicocconone-protein adduct, this ultrafast component remains almost unaffected but an additional rise time occurs, indicating the formation of a highly emissive species from the locally excited state. This phenomenon is central to the extraordinary applications of epicocconone in biotechnology. The magnitude of the rise time of the butylamine adduct is similar to that of the longer component of the decay of epicocconone in acetonitrile, suggesting that the dynamics of epicocconone and its butylamine adduct are similar. The ultrafast component is slowed upon increasing the viscosity of the solvent. This results in a marked increase in quantum yield and suggests that it corresponds to rapid bond isomerization, leading to a nonradiative decay. Surprisingly, in water/sucrose mixtures, the ultrafast component remains unaffected but there is still an increase in quantum yield, suggesting that there are at least two nonradiative pathways, one involving bond isomerization and another involving proton transfer. The correct interpretation of these data will allow the design of second generation protein stains based on the epicocconone scaffold with increased quantum yields and photostability. PMID:21827198

Chatterjee, Soumit; Burai, Tarak Nath; Karuso, Peter; Datta, Anindya

2011-09-15

443

Preoperative Iodine Staining May Complicate the Demarcation of Esophageal Carcinoma  

PubMed Central

A 53-year-old man was suspected of having an esophageal neoplasm. An endoscopic examination including Lugol chromoendoscopy suggested an esophageal squamous cell neoplasm limited to the lamina propria. A targeted biopsy showed atypical squamous cells, and an endoscopic submucosal dissection was performed 22 days after the previous endoscopy. Although a single 40 mm unstained area was observed by preoperative Lugol chromoendoscopy, intraoperative endoscopy revealed a 25 mm iodine-unstained area, with small unstained areas scattered on the oral side. We included the small unstained areas in the extent of the resection through assessment by preoperative endoscopy. Histopathologically, the tumor extent appeared to coincide with the preoperative assessment. Tumor cells were found in the basal-parabasal layers of the mucosa, in which small unstained areas were scattered, although the superficial layers exhibited well-differentiated cells containing glycogen in the cytoplasm. Although Lugol chromoendoscopy, which can induce chemical esophagitis, is widely used, re-epithelialization after mucosal damage by preoperative iodine staining may complicate the intraoperative demarcation of tumors.

Asada-Hirayama, Itsuko; Kodashima, Shinya; Niimi, Keiko; Mochizuki, Satoshi; Yamamichi, Nobutake; Fujishiro, Mitsuhiro; Matsusaka, Keisuke; Fukayama, Masashi; Koike, Kazuhiko

2013-01-01

444

Preoperative iodine staining may complicate the demarcation of esophageal carcinoma.  

PubMed

A 53-year-old man was suspected of having an esophageal neoplasm. An endoscopic examination including Lugol chromoendoscopy suggested an esophageal squamous cell neoplasm limited to the lamina propria. A targeted biopsy showed atypical squamous cells, and an endoscopic submucosal dissection was performed 22 days after the previous endoscopy. Although a single 40 mm unstained area was observed by preoperative Lugol chromoendoscopy, intraoperative endoscopy revealed a 25 mm iodine-unstained area, with small unstained areas scattered on the oral side. We included the small unstained areas in the extent of the resection through assessment by preoperative endoscopy. Histopathologically, the tumor extent appeared to coincide with the preoperative assessment. Tumor cells were found in the basal-parabasal layers of the mucosa, in which small unstained areas were scattered, although the superficial layers exhibited well-differentiated cells containing glycogen in the cytoplasm. Although Lugol chromoendoscopy, which can induce chemical esophagitis, is widely used, re-epithelialization after mucosal damage by preoperative iodine staining may complicate the intraoperative demarcation of tumors. PMID:23898393

Asada-Hirayama, Itsuko; Ono, Satoshi; Kodashima, Shinya; Niimi, Keiko; Mochizuki, Satoshi; Yamamichi, Nobutake; Fujishiro, Mitsuhiro; Matsusaka, Keisuke; Fukayama, Masashi; Koike, Kazuhiko

2013-07-01

445

A robust, efficient and flexible method for staining myelinated axons in blocks of brain tissue.  

PubMed

Previous studies have demonstrated the utility of the gold chloride method for en bloc staining of a bisected brain in mice and rats. The present study explores several variations in the method, assesses its reliability, and extends the limits of its application. We conclude that the method is very efficient, highly robust, sufficiently accurate for most purposes, and adaptable to many morphometric measures. We obtained acceptable staining of commissures in every brain, despite a wide variety of fixation methods. One-half could be stained 24 h after the brain was extracted and the other half could be stained months later. When staining failed because of an exhausted solution, the brain could be stained successfully in fresh solution. Relatively small changes were found in the sizes of commissures several weeks after initial fixation or staining. A half brain stained to reveal the mid-sagittal section could then be sectioned coronally and stained again in either gold chloride for myelin or cresyl violet for Nissl substance. Uncertainty, arising from pixelation of digitized images was far less than errors arising from human judgments about the histological limits of major commissures. Useful data for morphometric analysis were obtained by scanning the surface of a gold chloride stained block of brain with an inexpensive flatbed scanner. PMID:12606069

Wahlsten, Douglas; Colbourne, Frederick; Pleus, Richard

2003-03-15

446

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

PubMed Central

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

Woods, G L; Walker, D H

1996-01-01

447

Computerized prototypes of DAPI-stained chromosomes for FISH analysis  

SciTech Connect

DAPI is fluorescent dye widely used in chromosome counterstaining for fluorescence in situ hybridization (FISH) experiments. It produces a Q-banding pattern that allows chromosomes to be identified and permits molecular probes to be assigned to specific cytogenetic bands. Using a statistical procedure based on eigenanalysis, we have extracted features from digital images of DAPI-stained chromosomes and constructed prototypes of each of the 24 human chromosomes. The features of prototypes are directly proportional, in intensity profile and band location, to those of real chromosomes. The prototype`s intensity profile can be translated into cytogenetic bands to provide a computer-based strategy for chromosome mapping and analysis amenable to automation. Data have been obtained using images from the 24 human chromosomes and mouse X chromosome. Moreover, the same procedure is general and can be used for the analysis of chromosomes from other species, as well as with banding techniques other than those using DAPI. Images of hybridization patterns produced by complex probes are also suitable for this analysis. The speed and flexibility of the procedure opens the way to application so far unexplored, such as computer-assisted chromosome band assignment of probe; combined analysis of multiple, geometrically distorted chromosomes; and the direct comparison of raw data from different experiments. The applications will not be limited to mapping experiments but will include analysis of chromosome structure, variability and analysis of the pattern of chromosome distribution of repetitive sequences. The results from such analysis is suitable for objective statistical evaluation and, eventually, for autonomous machine interpretation.

Baldini, A.; Smith, L.C.; Knapp, R.D. [Baylor College of Medicine, Houston, TX (United States)

1994-09-01

448

Fluorescent Treponemal Antibody Absorption Double-Staining Test Evaluation  

PubMed Central

The fluorescent treponemal antibody absorption (FTA-ABS) double-staining (DS) test has been developed for microscopes equipped with incident illumination, and the procedure offers many advantages over the FTA-ABS test when tests are performed with this equipment. In this study, 346 fresh sera, including 35 from patients with syphilis, were evaluated by the FTA-ABS DS test. Parameters for investigation included two readers, each using a different microscope; a new FTA-ABS DS test reporting system; sera heated at 56C for 30 min versus unheated sera; and sera retested after at least 2 weeks of freezer storage. Agreement for FTA-ABS DS test readings between the two microscopes was 99%. Between-test agreement for the FTA-ABS test with the conventional reporting system and the FTA-ABS DS test with the new reporting system was 95%. Sensitivity calculations based on reactivity for the 35 syphilis sera were 94% for the FTA-ABS DS test and 91% for the FTA-ABS test. Specificity calculations based on non-reactivity of nonsyphilis sera were 98% for the FTA-ABS DS test and 93% for the FTA-ABS test. Differences in percentages appeared to be related to borderline readings in the FTA-ABS test. For example, if the same reporting system was used for the reference FTA-ABS test, the specificity was 97%. When sera were examined within 48 h, no difference was observed in results obtained with heated and unheated sera. Sera frozen for 2 weeks showed comparable results in the FTA-ABS DS test and the FTA-ABS test. These findings strongly support the recommendation that the FTA-ABS DS test be accepted as a confirmatory test for syphilis. The new reporting system for the FTA-ABS DS test would be advantageous for the reference FTA-ABS procedure.

Farshy, Carol E.; Kennedy, Edward J.; Hunter, Elizabeth F.; Larsen, Sandra A.

1983-01-01

449

High resolution structures of the 4-4-20 Fab-fluorescein complex in two solvent systems: effects of solvent on structure and antigen-binding affinity.  

PubMed Central

Three-dimensional structures were determined for three crystal forms of the antigen binding fragment (Fab) of anti-fluorescein antibody 4-4-20 in complex with fluorescein. These included 1) a triclinic (P1) form crystallized in 47% (v/v) 2-methyl-2,4-pentanediol (MPD); 2) a triclinic (P1) form crystallized in 16% (w/v) poly(ethylene glycol), molecular weight 3350 (PEG); and 3) a monoclinic (P21) form crystallized in 16% PEG. Solvent molecules were added to the three models and the structures were refined to their diffraction limits (1.75-A, 1.78-A, and 2.49-A resolution for the MPD, triclinic PEG, and monoclinic PEG forms, respectively). Comparisons of these structures were interesting because 4-4-20 exhibited a lower antigen-binding affinity in 47% MPD (Ka = 1.3 x 10(8) M-1) than in either 16% PEG (Ka = 2.9 x 10(9) M-1) or phosphate-buffered saline (Ka = 1.8 x 10(10) M-1). Even though the solution behavior of the antibody was significantly different in MPD and PEG, the crystal structures were remarkably similar. In all three structures, the fluorescein-combining site was an aromatic slot formed by tyrosines L32, H96, and H97 and tryptophans L96 and H33. In addition, several active site constituents formed an electrostatic network with the ligand. These included a salt link between arginine L34 and one of fluorescein's enolate oxygen atoms, a hydrogen bond between histidine L27d and the second enolic group, a hydrogen bond between tyrosine L32 and the phenylcarboxylate group, and two medium range (approximately 5 A) electrostatic interactions with lysine L50 and arginine H52. The only major difference between the triclinic MPD and PEG structures was the degree of hydration of the antigen-combining site. Three water molecules participated in the above electrostatic network in the MPD structure, while eight were involved in the PEG structure. Based on this observation, we believe that 4-4-20 exhibits a lower affinity in MPD due to the depletion of the hydration shell of the antigen-combining site. Images FIGURE 3 FIGURE 4 FIGURE 5 FIGURE 6 FIGURE 7

Herron, J N; Terry, A H; Johnston, S; He, X M; Guddat, L W; Voss, E W; Edmundson, A B

1994-01-01

450

[Specimen preparation for ABO determination of various kinds of blood stains].  

PubMed

Trying to optimize the preparation of blood stains, we found methanol fixation not to produce very good results for the determination of ABO blood group antigens. It is advantageous to transfer blood stains before testing to cotton cloth. This transfer is also of practical use if blood stains are to be saved on a smooth surface for lateral determination. We testet on 35 different carrier materials, on which blood stains in casework often were found, whether blood grouping gave better results on either the original material or after transfer. Results are shown on a table. The test revealed, that solubility of the stain in aqua dest is a good sign for a successful transfer. Blood stains on pine-wood soil, soil and loam were not suited for ABO grouping. PMID:2802924

Ltterle, J; Gloss, R; Bauer, I

1989-01-01

451

Chemical enhancement of footwear impressions in blood on fabric - part 1: protein stains.  

PubMed

A range of protein stains were utilised for the enhancement of footwear impressions on a variety of fabric types of different colours with blood as a contaminant. A semi-automated stamping device was used to deliver test impressions at a set force to minimise the variability between impressions; multiple impressions were produced and enhanced by each reagent to determine the repeatability of the enhancement. Results indicated that while most protein stains used in this study successfully enhanced impressions in blood on light coloured fabrics, background staining caused interference on natural fabrics. Enhancement on dark coloured fabrics was only achieved using fluorescent protein stains, as non-fluorescent protein stains provided poor contrast. A further comparison was performed with commercially available protein staining solutions and solutions prepared within the laboratory from the appropriate chemicals. Both solutions performed equally well, though it is recommended to use freshly prepared solutions whenever possible. PMID:21889106

Farrugia, Kevin J; Savage, Kathleen A; Bandey, Helen; Nic Daid, Niamh

2011-09-01

452