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1

Fluorescein diacetate as a viability stain for tree roots and seeds  

Microsoft Academic Search

Fluorescein diacetate (FDA) was tested as a viability stain for roots of green ash as well as for seeds of green ash and 10 other tree species. The viability level indicated by FDA staining of green ash roots agreed well with root growth potential results, bud condition assessment, and foliage browning measurements. In seed viability experiments, the FDA staining intensity

Thomas L. Noland; Gina H. Mohammed

1997-01-01

2

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...

3

ASSAY FOR FLUORESCEIN DIACETATE HYDROLYTIC ACTIVITY: OPTIMIZATION FOR SOIL SAMPLES  

Technology Transfer Automated Retrieval System (TEKTRAN)

With the increased interest in integrated soil bioecosystem studies, there has been a need to have a method of measuring overall microbial activity potential. Hydrolysis of fluorescein diacetate (3',6'-diacetylfluorescein [FDA]) has been suggested as a possible method because the ubiquitous lipase, ...

4

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

Microsoft Academic Search

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

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

2001-01-01

5

Microbial cell responses to a non-ionic surfactant. II. Effects as assessed by fluorescein diacetate uptake  

Microsoft Academic Search

Summary The effects of a non-ionic surfactant, Pluronic F-68, on uptake of fluorescein diacetate (FDA) into yeast cells as measured by increase in fluorescence atca. 500 nm have been studied. The rate of FDA uptake was increased almost 2.5 fold by incubating cells with up to 5% commercial grade pluronic but this depended on source and degree of purity. Dye

Alastair T. King; Kenneth C. Lowe; Bernard J. Mulligan

1988-01-01

6

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

PubMed Central

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

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

2012-01-01

7

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

8

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

NASA Astrophysics Data System (ADS)

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

Yamada, Kengo; Yoshida, Katsuyuki; Sonoda, Ryoichi

9

21 CFR 582.6197 - Calcium diacetate.  

Code of Federal Regulations, 2011 CFR

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

2011-04-01

10

21 CFR 182.6197 - Calcium diacetate.  

Code of Federal Regulations, 2010 CFR

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

2010-04-01

11

21 CFR 182.6197 - Calcium diacetate.  

Code of Federal Regulations, 2011 CFR

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

2011-04-01

12

21 CFR 582.6197 - Calcium diacetate.  

Code of Federal Regulations, 2010 CFR

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

2010-04-01

13

21 CFR 582.6197 - Calcium diacetate.  

Code of Federal Regulations, 2012 CFR

... 2012-04-01 2012-04-01 false Calcium diacetate. 582.6197 Section 582.6197...RECOGNIZED AS SAFE Sequestrants 2 § 582.6197 Calcium diacetate. (a) Product. Calcium diacetate. (b) Conditions of use....

2012-04-01

14

21 CFR 182.6197 - Calcium diacetate.  

Code of Federal Regulations, 2014 CFR

... 2014-04-01 2014-04-01 false Calcium diacetate. 182.6197 Section 182.6197...RECOGNIZED AS SAFE Sequestrants 1 § 182.6197 Calcium diacetate. (a) Product. Calcium diacetate. (b) Conditions of use....

2014-04-01

15

21 CFR 182.6197 - Calcium diacetate.  

Code of Federal Regulations, 2012 CFR

... 2012-04-01 2012-04-01 false Calcium diacetate. 182.6197 Section 182.6197...RECOGNIZED AS SAFE Sequestrants 1 § 182.6197 Calcium diacetate. (a) Product. Calcium diacetate. (b) Conditions of use....

2012-04-01

16

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

17

21 CFR 582.6197 - Calcium diacetate.  

Code of Federal Regulations, 2014 CFR

... 2014-04-01 2014-04-01 false Calcium diacetate. 582.6197 Section 582.6197...RECOGNIZED AS SAFE Sequestrants 2 § 582.6197 Calcium diacetate. (a) Product. Calcium diacetate. (b) Conditions of use....

2014-04-01

18

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

19

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

PubMed Central

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

2014-01-01

20

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. PMID:24709799

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

2013-01-01

21

Identification of senescence and death in Emiliania huxleyi and Thalassiosira pseudonana: Cell staining, chlorophyll alterations, and  

E-print Network

permeability, hydrolytic enzyme, and caspase-like activities using fluorescent cell stains to document diacetate (CMFDA; hydrolytic activity), whereas E. huxleyi developed two distinct CMFDA populations

Berges, John A.

22

The Cellular Basis for Biocide-Induced Fluorescein Hyperfluorescence in Mammalian Cell Culture  

PubMed Central

Clinical examination of the ocular surface is commonly carried out after application of sodium fluorescein in both veterinary and medical practice by assessing the resulting ‘staining’. Although localized intensely stained regions of the cornea frequently occur after exposure to ‘adverse’ clinical stimuli, the cell biology underlying this staining is unknown, including whether intense fluorescein staining indicates the presence of damaged cells. Ocular exposure to certain contact lens multipurpose solutions (MPS) gives rise to intense fluorescein staining referred to as solution induced corneal staining (SICS), and we have made use of this phenomenon with Vero and L929 cell culture models to investigate the fundamental biology of fluorescein interactions with cells. We found that all cells take up fluorescein, however a sub-population internalize much higher levels, giving rise to brightly staining ‘hyperfluorescent’ cells within the treated cultures, which contain fluorescein throughout the cell cytoplasm and nucleus. The numbers of these hyperfluorescent cells are significantly increased after exposure to MPS associated with SICS. Surprisingly, hyperfluorescent cells did not show higher levels of staining with propidium iodide, a marker of lysed cells. Consistently, treatment with the cytolytic toxin benzalkonium chloride resulted in almost all cells staining with propidium iodide, and the complete abolition of fluorescein hyperfluorescence. Finally we found that internalization of fluorescein and its loss from treated cells both require cellular activity, as both processes were halted after incubation at 4°C. We conclude that fluorescein hyperfluorescence can be replicated in three diverse cell cultures, and is increased by MPS-treatment, as occurs clinically. The process involves the concentration of fluorescein by a sub-population of cells that are active, and does not occur in lysed cells. Our data suggest that corneal staining in the clinic reflects active living cells, and is not directly caused by dead cells being produced in response to adverse clinical stimuli. PMID:24489650

Bakkar, May M.; Hardaker, Luke; March, Peter; Morgan, Philip B.; Maldonado-Codina, Carole; Dobson, Curtis B.

2014-01-01

23

The cellular basis for biocide-induced fluorescein hyperfluorescence in mammalian cell culture.  

PubMed

Clinical examination of the ocular surface is commonly carried out after application of sodium fluorescein in both veterinary and medical practice by assessing the resulting 'staining'. Although localized intensely stained regions of the cornea frequently occur after exposure to 'adverse' clinical stimuli, the cell biology underlying this staining is unknown, including whether intense fluorescein staining indicates the presence of damaged cells. Ocular exposure to certain contact lens multipurpose solutions (MPS) gives rise to intense fluorescein staining referred to as solution induced corneal staining (SICS), and we have made use of this phenomenon with Vero and L929 cell culture models to investigate the fundamental biology of fluorescein interactions with cells. We found that all cells take up fluorescein, however a sub-population internalize much higher levels, giving rise to brightly staining 'hyperfluorescent' cells within the treated cultures, which contain fluorescein throughout the cell cytoplasm and nucleus. The numbers of these hyperfluorescent cells are significantly increased after exposure to MPS associated with SICS. Surprisingly, hyperfluorescent cells did not show higher levels of staining with propidium iodide, a marker of lysed cells. Consistently, treatment with the cytolytic toxin benzalkonium chloride resulted in almost all cells staining with propidium iodide, and the complete abolition of fluorescein hyperfluorescence. Finally we found that internalization of fluorescein and its loss from treated cells both require cellular activity, as both processes were halted after incubation at 4 °C. We conclude that fluorescein hyperfluorescence can be replicated in three diverse cell cultures, and is increased by MPS-treatment, as occurs clinically. The process involves the concentration of fluorescein by a sub-population of cells that are active, and does not occur in lysed cells. Our data suggest that corneal staining in the clinic reflects active living cells, and is not directly caused by dead cells being produced in response to adverse clinical stimuli. PMID:24489650

Bakkar, May M; Hardaker, Luke; March, Peter; Morgan, Philip B; Maldonado-Codina, Carole; Dobson, Curtis B

2014-01-01

24

Gram stain  

MedlinePLUS

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

25

Photodynamic effect of argon and diode laser on cholesteatoma cell cultures after intravital staining with absorption enhancers.  

PubMed

Chronic epitympanic otitis media, or chronic suppurative osteitis, is a destructive form of chronic middle-ear inflammation. The therapy of choice is complete surgical removal of the squamous epithelium from the middle ear. It is often impossible to inspect all areas of the middle ear with the posterior canal wall intact. Not all recesses can be reliably monitored with the microscope, particularly in the area of the antrum and hypotympanum. Residual squamous epithelium here causes frequent recurrences following cholesteatoma surgery. This study examines the effect of argon and diode lasers on cholesteatoma tissue. The aim is to develop a laser treatment selectively directed against cholesteatoma cells that can be performed after cholesteatoma surgery to eliminate any residual squamous epithelium. Intraoperatively harvested monolayer-cultured cholesteatoma cells stained in vivo with various absorption enhancers served as the in vitro examination model. Argon (499 nm) and diode lasers (810 nm) were applied since their irradiation has an appropriate tissue penetration depth and is absorbed by various chromophores such as neutral red (475-500 nm), fluorescein (488 nm), and indocyanine green (790-810). Intracellular staining of cultured cells increased the optical density at the wavelength corresponding to the dye. Neutral red damaged 50-60% of cultured cells merely by intracellular accumulation at high concentrations. An additive cell destruction of about 30% was achieved by also applying argon laser irradiation. Fluorescein diacetate caused no appreciable stain-induced damage to cultured cholesteatoma cells. Argon laser irradiation destroyed up to 60% of the cultures. Indocyanine green resulted in only minor damage to cultured cells. The diode laser destroyed up to 60% of the irradiated cells. Selective staining of cholesteatoma cells was not achieved with any of the dyes examined. Thus, other stained tissue could be damaged. Staining and subsequent laser irradiation destroys up to 60% of cultured cholesteatoma cells. Unstained irradiated cells are not affected. Indocyanine green and fluorescein are nontoxic and may thus be used as absorption enhancers. The diode and argon lasers appear to be basically suitable. Cell staining is not selective, i.e., other tissues would also be stained and damaged. To avoid such unwanted damage, it would be desirable to couple the chromophore to a specific antibody that binds only to cholesteatoma cells. PMID:15772874

Sedlmaier, B; Franke, A; Sudhoff, H; Jovanovic, S; Haisch, A

2005-01-01

26

Combinatorial discovery of peptide dendrimer enzyme models hydrolyzing isobutyryl fluorescein.  

PubMed

Two 6750-membered one-bead-one-compound peptide dendrimer combinatorial libraries L (X(4))(8)(LysX(3))(4)(LysX(2))(2)LysX(1) (X(1-4) = 14 different amino acids or deletion, Lys = branching lysine residue) and AcL (with N-terminal acetylation) were prepared by split-and-mix solid phase peptide synthesis. Screening toward fluorogenic substrates for esterase and aldolase activities using the in silica off-bead assay (N. Maillard et al., J. Comb. Chem. 2009, 11, 667-675) and bead decoding by amino acid analysis revealed histidine containing sequences active against fluorescein diacetate. Isobutyryl fluorescein, a related hydrophobic fluorogenic substrate, was preferentially hydrolyzed by dendrimers from library AcL containing hydrophobic residues such as AcH3 (AcHis)(8)(LysLeu)(4)(LysVal)(2)LysLysOH, compared to simple oligohistidine peptides as reference catalysts. Polycationic dendrimers from library L with multiple free N-termini such as H8 (His)(8)(Lys?Ala)(4)(LysThr)(2)LysaProNH(2) (aPro = (2S,4S)-4-aminoproline) showed stronger reactivity toward 8-acetoxypyrene-1,3,6-trisulfonate with partial acylation of N-termini. These experiments highlight the role of noncatalytic amino acids to determine substrate selectivity in peptide dendrimer esterase models. PMID:21438622

Maillard, Noélie; Biswas, Rasomoy; Darbre, Tamis; Reymond, Jean-Louis

2011-05-01

27

Stain Reagent Reversible Stain Kits  

E-print Network

GelCode ® Blue Stain Reagent GelCode ® SilverSNAP TM Stain GelCode ® E-Zinc TM Reversible Stain ease of use and relatively good sensitivity. Although alternative methods, such as silver staining,1 as a colloidal suspension. The colloidal particles do not penetrate the gel, but the dye molecules are extracted

Lebendiker, Mario

28

Quantitative studies of immunofluorescent staining*  

PubMed Central

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

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

1968-01-01

29

FLUORESCEIN COLCHICINE Synthesis, Purification, and Biological Activity  

E-print Network

colchicine derivative permits the localization of colchicine-binding receptors in cells. Fluorescein), stimulation of antibody production (32), and modulation of surface receptor movements (36, 44). The presence (Sigma Chemical Co., St. Louis, Mo.) and deacetyl colchicine will be described in detail in Results

Clark, John

30

Clinical staining of the ocular surface: mechanisms and interpretations.  

PubMed

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

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

2015-01-01

31

Accidental intra-arterial injection of fluorescein dye.  

PubMed

During fluorescein angiography, sodium fluorescein dye intended for intravenous use was inadvertently injected into an artery in the antecubital fossa. An immediate and dramatic orange discoloration of the skin distal to the injection combined with intense burning pain of the right forearm and hand were noted. The patient was treated with ice packs and analgesics. The fluorescein angiogram showed a delayed arm to eye circulation time, but was of normal quality. There were no long-term complications. PMID:6521981

Bovino, J A; Marcus, D F

1984-12-01

32

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 ...

33

Fluorescent Sensors for Zn2+ Based on a Fluorescein Platform  

E-print Network

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

Tsien, Roger Y.

34

Coffee stain on textiles. Mechanisms of staining and stain removal  

Microsoft Academic Search

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

Erik Kissa

1995-01-01

35

Fluorescein-labeled glutathione to study protein S-glutathionylation.  

PubMed

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

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

2010-07-01

36

Acid-fast stain  

MedlinePLUS

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

37

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; ...

38

Stool Gram stain  

MedlinePLUS

... series of special stains are added to the sample. The lab team member looks at the stained smear under the microscope to check for bacteria. The color, size, and shape of the cells help identify the ...

39

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 ...

40

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

41

Cine photography and video recording of anterior segment fluorescein angiography  

Microsoft Academic Search

A description is given of apparatus and technique for carrying out cine photography and video recording of anterior segment fluorescein angiography. We found cine best for single-frame analysis and video tape recording less expensive.

R J Marsh; S M Ford

1978-01-01

42

Quick Stain Removal Guide  

E-print Network

. Over time, soil can build up on some fabrics, espe- cially polyesters. A pretreatment product may actually super-clean an area, and may resemble a bleached spot. You can usually correct this by treating the entire garment with a prespotter... or presoak and rewashing with extra detergent. Treat the stain quickly. Time and heat exposure make removing stains harder. Use a blotting motion. Work from the inside of the garment or back of the stain to force the stain out rather than into the fabric...

Brown, Pamela J.

1998-07-29

43

Anthralin stain removal.  

PubMed

Results of an anthralin stain removal study on white 65% polyester/35% cotton, white 100% polyester, white 100% cotton, a white shower curtain, white tile with crevice, and white ceramic shower tile are reported. An optimum stain removal technic was developed by using a 10-minute soak in full-strength chlorine bleach (Good Measure or Clorox) followed by a water rinse and air drying. This technic completely removed all stains of 24-hour duration from the test fabrics. The stain removal test on shower curtains, floor tiles, and ceramic shower tiles was also discussed. PMID:2431014

Wang, J C; Krazmien, R J; Dahlheim, C E; Patel, B

1986-11-01

44

Haematoporphyrin diacetate: a probe to distinguish malignant from normal tissue by selective fluorescence.  

PubMed Central

Haematoporphyrin diacetate has been synthesized and purified. Following introduction into mice bearing squamous-cell carcinomas it was found to produce a fluorescence of the tumour tissue which was much stronger than that of the surrounding normal tissue. Haematoporphyrin diacetate was also shown to be the major active component of the impure compound known as haematoporphyrin derivative which has been widely used in tumour fluorescence studies. Images Fig. 1 Fig. 2 Fig. 3 PMID:6775667

Henderson, R. W.; Christie, G. S.; Clezy, P. S.; Lineham, J.

1980-01-01

45

Field-scale evaluation of CFDA\\/SE staining coupled with multiple detection methods for assessing the transport of bacteria in situ  

Microsoft Academic Search

This research was undertaken to evaluate staining with the fluorescent compound CFDA\\/SE (5-(and-6-)-carboxyfluorescein diacetate, succinimidyl ester) coupled with multiple cell detection methodologies as a means to monitor bacterial transport during field-scale experiments. Stained cells of Comamonas sp. strain DA001 were injected into a shallow, aerobic aquifer in Oyster, VA, USA. Groundwater samples analyzed in the laboratory using epifluorescence microscopy, flow

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

2001-01-01

46

A New Fluorescence Staining Assay for Visualizing Living Microorganisms in Soil  

PubMed Central

5- (and 6-)Sulfofluorescein diacetate (SFDA), which is converted to a fluorescent product by intracellular esterase activity, was used to stain living microorganisms, including bacteria, Saccharomyces cerevisiae, and fungi, in soil. SFDA (1 mM) dissolved in ethyl alcohol was added to an intact soil sample, and the preparation was examined with an epifluorescence microscope. Bright single cells and colonies of live bacteria were observed without interference from the autofluorescence of soil minerals and detritus. Cultured Escherichia coli was killed through heat treatment; thus, SFDA was concluded to stain only living cells. Microbial colonies obtained from natural soils and various cultured strains were tested. It was found that 151 of 154 colonies from natural soils were stained and that hyphae and spores from 1 of 28 cultured microbial strains were not stained. The SFDA method was successfully used to visualize and count bacteria in soil samples from Mount Shiga in Japan. PMID:16535127

Tsuji, T.; Kawasaki, Y.; Takeshima, S.; Sekiya, T.; Tanaka, S.

1995-01-01

47

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

48

Port-Wine Stains  

MedlinePLUS

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

49

Synthesis of fluorescein labeled 7-methylguanosinemonophosphate Amarnath Natarajan,a  

E-print Network

(an RNA helicase) and eIF4G (an adaptor pro- tein).2 The critical role played by translation a fluorescein labeled 7-methylguanosinemonophosphate cap ana- logue (Flu-cap, Fig. 1). When mixed with purified as a dimethylformamidine and the 3,4-dihydroxy as a di- methylacetonide, to generate the soluble and fully pro- tected

50

Selective perborate signaling by deprotection of fluorescein and resorufin acetates.  

PubMed

The acetate derivatives of fluorescein and resorufin exhibited a prominent turn-on type signaling behavior toward BO(3)(-) ions over other common anions. Signaling is based on the selective deprotection of acetate groups by perborate, which resulted in significant chromogenic and fluorogenic signaling. Compound 1 also exhibited a pronounced perborate selectivity over other commonly used oxidants in 90% aqueous acetonitrile solution. PMID:20222739

Choi, Myung Gil; Cha, Sunyoung; Park, Ji Eun; Lee, Haekyung; Jeon, Hye Lim; Chang, Suk-Kyu

2010-04-01

51

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

52

Lineage labeling of zebrafish cells with laser uncagable fluorescein dextran.  

PubMed

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

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

2011-01-01

53

Presurgical antiseptic efficacy of chlorhexidine diacetate and providone-iodine in the canine preputial cavity.  

PubMed

Antiseptic flushing of the canine prepuce and its exclusion from the surgical field are recommended before abdominal surgery to reduce the risk of bacterial contamination. The authors cultured the preputial cavity of 60 dogs prior to and following flushing with 0.05% chlorhexidine diacetate, 1% povidone-iodine, or 0.9% saline control. Bacterial growth was evaluated using a semiquantitative method, and bacterial organisms were subsequently identified. There were no significant differences between povidone-iodine and the saline control in any of the variables assessed. Chlorhexidine resulted in a significant decrease in the proportion of positive postflush cultures compared with povidone-iodine. Although not significant, the difference in adverse reactions between povidone-iodine (25%) and chlorhexidine diacetate (5%) suggests clinical relevance. Based on the results of this study, a 2 min flush with 0.05% chlorhexidine diacetate is recommended for presurgical preparation of the preputial cavity. PMID:22058347

Neihaus, Steven A; Hathcock, Terri L; Boothe, Dawn M; Goring, Robert L

2011-01-01

54

Ruby Protein Stains Instruction Manual  

E-print Network

SYPRO® Ruby Protein Stains Instruction Manual SYPRO® Ruby protein gel stain Catalog Numbers 170 ......................................................................................7 2.2 Visualizing and Imaging a Blot .................................................9 PART 4

Lebendiker, Mario

55

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

NASA Astrophysics Data System (ADS)

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

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

2011-04-01

56

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

57

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

58

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

Technology Transfer Automated Retrieval System (TEKTRAN)

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

59

Gram stain of skin lesion  

MedlinePLUS

... of different colored stains is applied to the sample. A laboratory team member examines the stained slide under a microscope, checking for bacteria. The color, size, and shape of the cells help identify the ...

60

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

61

Syntheses of (-)-cryptocaryolone and (-)-cryptocaryolone diacetate via a diastereoselective oxy-Michael addition and oxocarbenium allylation.  

PubMed

The total syntheses of both (-)-cryptocaryolone and (-)-cryptocaryolone diacetate is presented herein. The usage of a diastereoselective oxy-Michael addition/benzylidene acetal formation coupled with a selective axial oxocarbenium allylation allowed for the preparation of the ?-C-glycoside moiety present in the bicyclic bridged structure. In addition, the syn-1,3-diol of the linear portion was installed via a Wacker oxidation followed by a subsequent directed reduction of the appropriate homoallylic alcohol precursor. PMID:22827503

Albury, Aymara M M; Jennings, Michael P

2012-08-17

62

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

PubMed Central

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

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

1984-01-01

63

Chromoendoscopy and intravital staining techniques.  

PubMed

Chromoendoscopy and intravital staining techniques are synonymous methods for the endoscopic early detection of malignant changes in the intestinal tract. Endoscopic intravital staining involves the use of absorptive stains (methylene blue and Lugol's solution), contrast stains (indigo carmine) and reactive stains (Congo red). Lugol's iodine solution is used to identify superficial carcinomas in the squamous epithelium of the oesophagus. Methylene blue stains the specialized intestinal epithelium in Barrett's oesophagus and, in addition to this, is helpful in the diagnosis of dysplasia. Intravital staining with indigo carmine contributes to contrasting and accentuating changed mucosal processes. Together with Cresyl violet, contrast staining is particularly important in detecting small, early malignant changes in the colon. The use of chromoendoscopy enables a biopsy diagnosis of superficial dysplastic changes and an accurate delineation of carcinomatous areas. In conjunction with the modern video-endoscopy (high-resolution endoscopy and magnification endoscopy), vital staining forms the diagnostic foundation for the detection of early malignant changes in the gastrointestinal tract. These techniques are therefore prerequisites to local endoscopic tumour therapy (mucosectomy). Despite their increasing acceptance, these methods must prove their diagnostic merit in randomized studies. PMID:11030630

Jung, M; Kiesslich, R

1999-04-01

64

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

PubMed Central

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

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

2000-01-01

65

Development of a vital fluorescent staining method for monitoring bacterial transport in subsurface environments.  

PubMed

Previous bacterial transport studies have utilized fluorophores which have been shown to adversely affect the physiology of stained cells. This research was undertaken to identify alternative fluorescent stains that do not adversely affect the transport or viability of bacteria. Initial work was performed with a groundwater isolate, Comamonas sp. strain DA001. Potential compounds were first screened to determine staining efficiencies and adverse side effects. 5-(And 6-)-carboxyfluorescein diacetate, succinimidyl ester (CFDA/SE) efficiently stained DA001 without causing undesirable effects on cell adhesion or viability. Members of many other gram-negative and gram-positive bacterial genera were also effectively stained with CFDA/SE. More than 95% of CFDA/SE-stained Comamonas sp. strain DA001 cells incubated in artificial groundwater (under no-growth conditions) remained fluorescent for at least 28 days as determined by epifluorescent microscopy and flow cytometry. No differences in the survival and culturability of CFDA/SE-stained and unstained DA001 cells in groundwater or saturated sediment microcosms were detected. The bright, yellow-green cells were readily distinguished from autofluorescing sediment particles by epifluorescence microscopy. A high throughput method using microplate spectrofluorometry was developed, which had a detection limit of mid-10(5) CFDA-stained cells/ml; the detection limit for flow cytometry was on the order of 1,000 cells/ml. The results of laboratory-scale bacterial transport experiments performed with intact sediment cores and nondividing DA001 cells revealed good agreement between the aqueous cell concentrations determined by the microplate assay and those determined by other enumeration methods. This research indicates that CFDA/SE is very efficient for labeling cells for bacterial transport experiments and that it may be useful for other microbial ecology research as well. PMID:11010903

Fuller, M E; Streger, S H; Rothmel, R K; Mailloux, B J; Hall, J A; Onstott, T C; Fredrickson, J K; Balkwill, D L; DeFlaun, M F

2000-10-01

66

Automated Feulgen staining with a temperature-controlled staining machine.  

PubMed

A Shandon Varistain 24-3 staining machine was modified in order to run automated DNA Feulgen staining. Initial studies showed a strict dependence of the staining intensity (integrated optical density [IOD]) on the temperature of the DNA hydrolysis in 4 N HCl: a difference of 0.5 degrees C around the optimum hydrolysis temperature of 27.5 degrees C resulted in IOD differences of up to 7.8% in epithelial cells and up to 12.0% in lymphocytes. A temperature-controlled stainless steel cuvette, covered with a 4 N HCl-resistant material, was developed and integrated into the machine. Temperature measurements were performed at different positions in the cuvette and on glass slides with copper-constantan electrodes fixed on them; no temperature gradient could be detected within the cuvette. The adjusted temperature of 27.5 degrees C remained constant over 24 hours. The coefficient of variation (CV) of the staining intensity in lymphocytes between different areas on the same slide and between different slides of the same staining cycle was less than 0.6%. The CV between different staining cycles was 5.9%. This system for automated Feulgen staining thus gives reproducible and reliable results and may be introduced into routine diagnostic procedures. PMID:2472804

Chatelain, R; Willms, A; Biesterfeld, S; Auffermann, W; Böcking, A

1989-06-01

67

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. PMID:19815735

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

2010-01-01

68

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

PubMed Central

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

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

2014-01-01

69

Systematic study of fluorescein-functionalized macrophotoinitiators for colorimetric bioassays.  

PubMed

We report a systematic investigation of a set of photoreducible macrophotoinitiators for use in polymerization-based signal amplification. To test the dependence of photopolymerization responses on the number of photoinitiators localized per molecular recognition event, we gradually increased the number of photoinitiator molecules coupled to a constant scaffold macromolecule from an average of 7 per polymer to an average of 168 per polymer. To evaluate the capacity of the macrophotoinitiators to detect molecular recognition, we coupled neutravidin to these molecules to recognize biotin-labeled DNA immobilized on biochip test surfaces. Fluorescein macroinitiators were found to be useful in detecting molecular recognition above a threshold number of initiators per polymer. Above this threshold, increasing the number of initiators per macroinitiator resulted in increased signal strength. These findings demonstrate the feasibility of increasing the number of photoreducible initiators per binding event beyond three, the number used in previous studies, that the initiation reaction remains limiting in the range we investigated, and that the number of initiators per binding event in this system has a clear impact on assay sensitivity and signal strength. PMID:22404188

Lee, Jungkyu K; Heimer, Brandon W; Sikes, Hadley D

2012-04-01

70

[Chemical modification of steroid-hydroxylating monooxygenases with fluorescein isothiocyanate].  

PubMed

Chemical modifications of cytochrome P-450scc and cytochrome P-450(11) beta with fluorescein-, diiodofluorescein-, eosine- and rhodamine isothiocyanate have been carried out. At a low reagent/protein ratio and neutral pH, a selective chemical modification was known to take place which did not affect the spectral properties of cytochrome P-450scc. Covalent chromatography was found useful to discriminate between covalent modification of cytochrome P-450scc and non-specific binding of FITC with cytochrome P-450scc. Proteolytic modification of cytochrome P-450scc and structural analysis indicate that a lysine residue of the C-terminal sequence of cytochrome P-450scc is accessible to FITC. The residue was shown, by the analysis of the chymotryptic hydrolysate of the fragment F2, to be Lys338. Effect of modification with FITC on the interaction of cytochrome P-450scc with cholesterol or adrenodoxin, on the reduction kinetics and on the conversion of cholesterol to pregnenolone was also studied. PMID:1793433

Lapko, A G; Smettan, G; Rukpaul', K; Usanov, S A

1991-07-01

71

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

72

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

73

Development of resistance to chlorhexidine diacetate in Pseudomonas aeruginosa and the effect of a "residual" concentration.  

PubMed

Stable resistance in Pseudomonas aeruginosa NCIMB 10421 was obtained by step-wise exposure to gradually increasing concentrations of chlorhexidine diacetate (CHX). Repeated exposure to a proposed "residual" (sub-MIC) concentration of CHX also created stable resistance. Resistance was also developed by a single exposure to the "residual" concentration of CHX, but this was unstable. Similar experiments with Escherichia coli and CHX or cetylpyridinium chloride resulted in no significant increase in resistance. Antibiotic susceptibility profiles of the CHX-resistant P. aeruginosa cultures showed no cross-resistance, although some of the cultures were resistant to benzalkonium chloride. PMID:11170761

Thomas, L; Maillard, J Y; Lambert, R J; Russell, A D

2000-12-01

74

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

2014-01-01

75

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

76

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

EPA Science Inventory

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

77

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

E-print Network

it is dependent on pH, having a lactone-locked formed under acidic conditions thus effectively inhibiting its fluorescent properties. Fluorescein does not transform from an open form to a lactone form directly. Instead, it consists of five different... Mannich conditions. The synthesis call for the substitution of resorcinol with 2-methylresorcinol in the standard fluorescein condensation procedure, followed by protection of the phenolic groups in the lactone form of the dye. The next step involves...

Castro, Juan C.

2010-07-14

78

The growth of thin fluorescein films on Ag (1 1 0)  

NASA Astrophysics Data System (ADS)

The thin fluorescein films grown on Ag (1 1 0) have been studied by the ultraviolet photoemission spectroscopy (UPS). Four emission features are located at 2.7, 3.8, 7.4 and 9.8 eV below the Fermi level, respectively. The angle-resolved ultraviolet photoemission spectroscopy (ARUPS) measurements show that the Triring plane of fluorescein is nearly parallel to the substrate and the axis of C dbnd O is close to the [1 1¯ 0] azimuth.

Qian, H. Q.; Mao, H. Y.; Chen, Q.; Song, F.; Hu, Y. W.; Huang, H.; Zhang, H. J.; Li, H. Y.; He, P. M.; Bao, S. N.

2006-12-01

79

Study of Choroidal Blood Flow by Comparison of SLO Fluorescein Angiography and Microspheres  

Microsoft Academic Search

Choroidal hemodynamics estimated with parameters describing the dye build-up curves obtained with video fluorescein angiography, were compared with a classical regional blood flow measurement: radioactively labelled microspheres.Video fluorescein angiograms (Rodenstock's SLO 101) and microspheres blood flow measurements were made in 13 anaesthetized pigmented rabbits. Ocular perfusion pressures were varied from 60 to 15 mmHg by changing the intraocular pressure. The

H. FERDINAND A. DUIJM; ALEXANDER H. F. RULO; MARIA ASTIN; OLAV MÄEPEA; THOMAS J. T. P. VAN DEN BERG; ERIK L. GREVE

1996-01-01

80

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

NASA Astrophysics Data System (ADS)

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

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

1994-06-01

81

Automated single-slide staining system  

NASA Technical Reports Server (NTRS)

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

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

1974-01-01

82

Papanicolaou staining--a review.  

PubMed

Some technical aspects of Papanicolaou staining are reviewed. The history of the technique is traced from Mallory's aniline blue technique through Masson's trichrome procedure to the techniques of Shorr. The histochemistry of the three Papanicolaou staining solutions (aluminum-"hematoxylin", OG and EA) is discussed. The structure of the aluminum-hematein chelate and its mode of action are considered. A tentative mechanism is proposed for the characteristic differential counterstaining produced by Papanicolaou techniques, i.e. orangeophilia vs cyanophilia. It is suggested that differential counterstaining occurs as a consequence of 1) differences in cytoplasmic density, 2) differences in the molecular size of the anionic dyes, and 3) the inhibitory effects of phosphotungstic acid on the binding of small dyes. The review considers some recent quantitative studies of Papanicolaou stained cells and outlines some modifications of Papanicolaou's procedures. The text concludes with a discussion of alternatives to the Papanicolaou technique. PMID:6195507

Marshall, P N

1983-05-01

83

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

Federal Register 2010, 2011, 2012, 2013, 2014

...estradiol; ethynodiol diacetate) Tablet, 0.035 mg; 1 mg. NDA 019190...levonorgestrel) , Inc., P.O. Tablet, 0.03 mg, 0.04 Box 8299...estradiol; levonorgestrel) Tablet, 0.03 mg, 0.04 mg, 0...FR Doc. 2011-31146 Filed 12-2-11; 8:45 am]...

2011-12-05

84

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

Technology Transfer Automated Retrieval System (TEKTRAN)

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

85

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

PubMed

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

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

2014-12-01

86

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

87

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

88

Whole Blood Cell Staining Device  

NASA Technical Reports Server (NTRS)

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

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

2000-01-01

89

Fluorogenic Substrate Detection of Viable Intracellular and Extracellular Pathogenic Protozoa  

NASA Astrophysics Data System (ADS)

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

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

1985-01-01

90

Methods for chromosome-specific staining  

DOEpatents

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

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

1995-01-01

91

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

PubMed

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

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

92

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

PubMed

Cytometric sorting of frozen-thawed sperm can overcome difficulties caused by the unavailability of sorting facilities on farms where semen is collected from male livestock. In order to optimize the cytometric sex-sorting procedure, effects of Hoechst33342 staining on the viability and cytometric sorting efficiency of frozen-thawed ram sperm were evaluated. The frozen-thawed sperm were stained with Hoechst33342 at various dye concentrations (80?M, 120?M, 160?M, 200?M, 240?M, or 320?M) for 45min to evaluate effects of dye dose. The frozen-thawed sperm were stained with 160?M Hoechst33342 for various durations (0min, 15min, 30min, 45min, 60min, 75min, or 90min) to evaluate effects of staining duration. Sperm motility and moving velocity were analyzed using a computer-assisted sperm analysis system (CASAS). Acrosome status, membrane integrity, and distribution of phosphatidylserine (PS) in Hoechst33342-stained sperm were analyzed using flow cytometry after staining with fluorescein isothiocyanate-labeled lectin from pisum sativum (FITC-PSA), Annexin V, or propidium iodide (PI). The fertility of Hoechst33342-stained sperm was analyzed by in vitro fertilization (IVF). A high-speed cell sorter was used to evaluate effects of Hoechst33342 staining on cytometric sex-sorting of frozen-thawed sperm. The motility, moving velocity, membrane integrity, and PS distribution of Hoechst33342-stained sperm were significantly different from that of immediately thawed sperm (P<0.05). However, there is no significant difference existing among the Hoechst33342-stained groups with respect to the above evaluated parameters. Additionally, along with the staining durations, the adverse effects of the staining procedure on sperm showed a steady increase. However, Hoechst33342 staining did not damage acrosome and in vitro fertilizing capability of frozen-thawed ram sperm. Results of cytometric sorting indicated that frozen-thawed sperm can be efficiently sorted into two sperm populations with X and Y chromosome when the Hoechst33342 concentration was 160?M. Moreover, when the staining duration was equal to or longer than 45min, the frozen-thawed sperm can be successfully sorted in the presence of 160?M Hoechst33342. In conclusion, Hoechst33342 staining can detrimentally influence viability of frozen-thawed ram sperm except acrosome and in vitro fertilizing capability. Accordingly, the minimum values of Hoechst33342 concentration and staining duration can be set at 160?M and 45min respectively. However, the maximum values of Hoechst33342 concentration and staining duration were not determined based on the current study. Further research on how to reduce injuries caused by freezing, thawing, and Hoechst33342 staining on frozen-thawed ram sperm is needed. PMID:25481668

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

2015-02-01

93

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

PubMed Central

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

Slifkin, M; Cumbie, R

1989-01-01

94

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

NASA Astrophysics Data System (ADS)

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

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

2013-01-01

95

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

Microsoft Academic Search

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

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

2010-01-01

96

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

PubMed Central

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

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

2012-01-01

97

The fluorescein-derived dye aminophenyl fluorescein is a suitable tool to detect hypobromous acid (HOBr)-producing activity in eosinophils.  

PubMed

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

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

2012-08-10

98

[Based on blood vessel edge feature fundus fluorescein angiography image splicing].  

PubMed

According to fundus fluorescein angiography images characteristics, this paper proposes a feature based image mosaic vessel edge method. First, detect edge of blood vessels by carrying on the pretreatment to the fundus fluorescein angiography image in the foundation, wavelet edge detection algorithm. Then, the matching method based on chain code feature is described. Finally, a local area based on gray level information of the image fusion method is applied to angiographic image series. Data processing results show that the method can generate an ideal mosaic effect. PMID:21954575

Cui, Dong; Liu, Minmin; Guo, Yongxin; Jiao, Qing

2011-05-01

99

The use of lysozyme modified with fluorescein for the detection of Gram-positive bacteria.  

PubMed

Lysozyme (1,4-?-N-acetylmuramidase) is commonly applied in the food, medical, and pharmaceutical industries. In this study, we tested a novel application of fluorescein-modified lysozyme (using carboxyfluorescein with a triazine-based coupling reagent) as a new tool for the detection of Gram-positive soil bacteria. The results, obtained by cultivation methods, fluorescence analysis, and laser interferometry, showed that, after optimization, fluorescein-modified lysozyme could be used to evaluate the prevalence of Gram-positive bacteria essential in bioremediation of soils with low pH, such as those degraded by sulfur. PMID:24916601

Arabski, Micha?; Konieczna, Iwona; Tusi?ska, Ewa; W?sik, S?awomir; Relich, Inga; Zaj?c, Krzysztof; Kami?ski, Zbigniew J; Kaca, Wies?aw

2015-01-01

100

Photodissociation of cold fluorescein monoanion studied using an electrostatic storage ring  

NASA Astrophysics Data System (ADS)

Photodissociation of fluorescein monoanions was studied by using an electrostatic storage ring. The ions are stored in an ion trap and the storage ring before laser irradiation. Photodissociation neutral spectra of the relaxed ions as a function of time consist of components with short and long lifetimes. By comparing their wavelength spectra with those in the liquid phase, it is concluded that the spectra originate from different tautomers of fluorescein monoanions. Moreover, via storage in the storage ring, one of them disappears due to interconversion.

Tanabe, Tetsumi; Saito, Manabu; Noda, Koji

2012-11-01

101

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

102

Investigation on interaction and sonodynamic damage of fluorescein derivants to bovine serum albumin (BSA) under ultrasonic irradiation  

NASA Astrophysics Data System (ADS)

The fluorescein derivants (Fluorescein: (2-(6-Hydroxy-3-oxo-(3H)-xanthen-9-yl) benzoic acid), Fluorescein-DA: (Bis [N,N-bis (carboxymethyl) aminomethyl] fluorescein) and Fluorescein-DAsbnd Fe(III): (Bis [N,N-bis (carboxymethyl) aminomethyl] fluoresceinsbnd Ferrous(III)) with a tricyclic plane structure were used to study the interaction and sonodynamic damage to bovine serum albumin (BSA) under ultrasonic irradiation through fluorospectrometry and UV-vis spectrophotometry. Besides, because of the existence of Fe(III) ion in Fluorescein-DAsbnd Fe(III), under ultrasonic irradiation the sonocatalytic activity in the damage of BSA molecules was also found. Three-dimensional fluorescence spectra and three-dimensional fluorescence contour profile spectra were mentioned to determine the fluorescence quenching and the conformation change of BSA in the absence and presence of these fluorescein derivants. As judged from the experimental results, the fluorescence quenching of BSA in aqueous solution caused by these fluorescein derivants were all attributed to static quenching process. The damage degree and mode were related to some factors such as ultrasonic irradiation time, fluorescein derivant concentration and ionic strength. Finally, several quenchers were used to determine the amount and kind of generated reactive oxygen species (ROS) during sonodynamic and sonocatalytic reaction processes. It suggests that these fluorescein derivants induce protein damage via various ROS, at least, including singlet oxygen (1O2) and hydroxyl radicals (rad OH). Perhaps, this paper may offer some important subjects for broadening the application of these fluorescein derivants in sonodynamic therapy (SDT) and sonocatalytic therapy (SCT) technologies for tumor treatment.

Zou, Mingming; Zhang, Lei; Wang, Jun; Wang, Qi; Gao, Jingqun; Fan, Ping

2013-06-01

103

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

104

Removal of Fluorescein using different iron oxides as adsorbents: effect of pH.  

PubMed

In this work the adsorption process of Fluorescein (dye with aril-methane group) as a function of pH on three different adsorbents: goethite, Co-goethite, and magnetite has been studied experimentally and theoretically. FTIR and Raman spectroscopy have been performed in an attempt to confirm the structure of surface complexes formed by sorption of the Fluorescein to different iron oxides. Typical anionic adsorption behaviour was observed for this dye onto goethite and Co-goethite whereas the adsorption level was practically constant in the range of pH studied when the adsorbent was magnetite. The diffuse layer model was employed to fit the experimental results. The surface complexes proposed from the adsorption data were in agreement with the patterns obtained from FTIR and Raman spectroscopy. The surface structure of the oxides affects the adsorption process and the final adsorbed amount at the equilibrium. Our model of diffuse double layer with the addendum of the effect of hydrophobic forces fits well the adsorption data of Fluorescein on iron oxides at different pH in the studied range. At lower pH electrostatic forces by ligand-exchange are predominant. In the range of pH 9-11 hydrophobic forces are managing the Fluorescein adsorption on the iron oxides, with the formation of outer-sphere complexes through van der Waals/hydrophobic forces. It is interesting that in the three iron oxides studied, the adsorbed amount in this range is similar. PMID:18308623

Pirillo, Silvina; Cornaglia, Laura; Ferreira, María Luján; Rueda, Elsa H

2008-11-15

105

Sweat pore mapping using a fluorescein-polymer composite film for fingerprint analysis.  

PubMed

A simple but efficient sweat pore mapping method based on a fluorescein-PVP composite film was developed for fingerprint analysis. The composite film displays a fluorometric turn-on response upon contact with a small quantity of water secreted from human sweat pores, allowing precise mapping of sweat pores on a fingertip. PMID:25604679

Pyo, Minkyeong; Lee, Joosub; Baek, Woohyun; Lee, Chan Woo; Park, Bum Jun; Kim, Jong-Man

2015-02-01

106

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

107

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

108

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

PubMed Central

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

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

2014-01-01

109

New developments in the filter test system for cytotoxicity testing  

Microsoft Academic Search

The objective of this study was (1) to improve the succinate dehydrogenase (SDH) staining procedure of the filter test system, and (2) to study the suitability of hydrolases as markers for cell vitality by means of fluorescein diacetate, instead of SDH. The test materials included zinc phosphate cements, conventional and light-cured glass ionomer cements, a composite resin, and methylmethacrylate monomer.

G. Schmalz; K.-A. Hiller; F. Dörter-Aslan

1994-01-01

110

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

EPA Science Inventory

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

111

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. ...

112

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

113

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

114

Rapid assessment of bacterial viability by flow cytometry  

Microsoft Academic Search

The ability of a flow cytometer to rapidly assess microbial viability was investigated using three vital stains: rhodamine 123 (Rh123); 3,3'-dihexyloxacarbocyanine iodide [DiOC6(3)] and fluorescein diacetate (FDA). Rh123 was found to clearly differentiate viable from non-viable bacteria. The methodology for staining bacteria with this dye was optimised. Rh123 was shown to stain and discriminate several different species of viable bacteria

J. P. Diaper; K. Tither; C. Edwards

1992-01-01

115

A lysine residue involved in the inhibition of vacuolar H +-pyrophosphatase by fluorescein 5?-isothiocyanate  

Microsoft Academic Search

Vacuolar proton pumping pyrophosphatase (H+-PPase; EC 3.6.1.1) plays a central role in the electrogenic translocation of protons from cytosol to the vacuole lumen at the expense of PPi hydrolysis. A fluorescent probe, fluorescein 5?-isothiocyanate (FITC), was used to modify a lysine residue of vacuolar H+-PPase. The enzymatic activity and its associated H+ translocation of vacuolar H+-PPase were markedly decreased by

Su Jing Yang; Shih Sheng Jiang; Ru Chuan Van; Yi Yuong Hsiao; Rong-Long Pan

2000-01-01

116

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

Microsoft Academic Search

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

Katja Berginc; Simon Žakelj; Lea Levstik; Darko Urši?; Albin Kristl

2007-01-01

117

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

PubMed Central

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

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

2014-01-01

118

A Method for Assessment of Blood Volume Parameters in Pregnant Sheep using Fluorescein-labelled Dextran  

Microsoft Academic Search

The assessment of blood volume parameters in clinical and research settings has been limited by methods that involve radioactivity, complex assays or are unreliable. We aimed to design a method for measuring blood volume parameters that was non-radioactive, simple, cheap and reliable. We have used a commercially available fluorescein-labelled 250kDa dextran, a large inert molecule, and have measured dilution of

C. W. H. Rumball; P. Van Zijl; M. D. Rutland; F. H. Bloomfield; J. E. Harding

2008-01-01

119

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

E-print Network

, hydrochloride (Molecular Probes E-2247), store at ­20 °C - strains of interest - PBS METHODS 1) Make a 10 mg/ml (10X) fluorescein aminomethyl stock in PBS in its opaque bottle (to protect it from light). It will be orange and very granular. This can be stored at 4 °C. 2) Remove 11 µl of stock and dilute in 99 µl PBS

Doering, Tamara

120

Use of the gram stain in microbiology.  

PubMed

The Gram stain differentiates bacteria into two fundamental varieties of cells. Bacteria that retain the initial crystal violet stain (purple) are said to be "gram-positive," whereas those that are decolorized and stain red with carbol fuchsin (or safranin) are said to be "gram-negative." This staining response is based on the chemical and structural makeup of the cell walls of both varieties of bacteria. Gram-positives have a thick, relatively impermeable wall that resists decolorization and is composed of peptidoglycan and secondary polymers. Gram-negatives have a thin peptidoglycan layer plus an overlying lipid-protein bilayer known as the outer membrane, which can be disrupted by decolorization. Some bacteria have walls of intermediate structure and, although they are officially classified as gram-positives because of their linage, they stain in a variable manner. One prokaryote domain, the Archaea, have such variability of wall structure that the Gram stain is not a useful differentiating tool. PMID:11475313

Beveridge, T J

2001-05-01

121

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

PubMed

A novel fluorescein derivative furfuraldehyde fluorescein hydrazone (FFH) has been synthesized by reacting fluorescein hydrazide with furfuraldehyde and characterized by (1)H NMR, (13)C NMR, MS and elemental analysis. Addition of Cu(2+) to the solution of FFH results in a rapid color change from colorless to yellow together with an obvious new band appeared at 502 nm in the UV-vis absorption spectra. This change is attributed to the spirocycle form of FFH opened via coordination with Cu(2+) in a 1:1 stoichiometry and their association constant is determined as 6.1×10(4) L mol(-1). Experimental results indicate that the FFH can provide a rapid, selective and sensitive response to Cu(2+) with a linear dynamic range 6.6-330 ?mol/L. Common interferent ions do not show any interference on the Cu(2+) determination. It is anticipated that FFH can be a good candidate probe and has potential application for Cu(2+) determination in aqueous solution. PMID:24370938

Zhang, Jiangang; Zhang, Li; Wei, Yanli; Ma, Jun; Shuang, Shaomin; Cai, Zongwei; Dong, Chuan

2014-03-25

122

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

NASA Astrophysics Data System (ADS)

A novel fluorescein derivative furfuraldehyde fluorescein hydrazone (FFH) has been synthesized by reacting fluorescein hydrazide with furfuraldehyde and characterized by 1H NMR, 13C NMR, MS and elemental analysis. Addition of Cu2+ to the solution of FFH results in a rapid color change from colorless to yellow together with an obvious new band appeared at 502 nm in the UV-vis absorption spectra. This change is attributed to the spirocycle form of FFH opened via coordination with Cu2+ in a 1:1 stoichiometry and their association constant is determined as 6.1 × 104 L mol-1. Experimental results indicate that the FFH can provide a rapid, selective and sensitive response to Cu2+ with a linear dynamic range 6.6-330 ?mol/L. Common interferent ions do not show any interference on the Cu2+ determination. It is anticipated that FFH can be a good candidate probe and has potential application for Cu2+ determination in aqueous solution.

Zhang, Jiangang; Zhang, Li; Wei, Yanli; Ma, Jun; Shuang, Shaomin; Cai, Zongwei; Dong, Chuan

2014-03-01

123

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

124

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

125

Pyrolytical characterization of transition metal complexes of cobalt, nickel, copper and zinc with ethylenediamine- N,N ?-diacetate  

Microsoft Academic Search

A series of octahedral complexes, [M(EDDA)(H2O)2] · H2O (where, M+2 = Co(II), Cu(II), Ni(II) and Zn(II); EDDA, ethylenediamine-N,N?-diacetate), was prepared and studied by means of thermogravimetry (TG) and differential thermal analysis (DTA). Their compositions\\u000a were investigated by elemental analysis in order to ensure their purity and structural elucidation was based on spectral and\\u000a magnetic properties. Thermal decomposition of these distorted octahedral complexes, [Ni(EDDA)(H2O)2], [Co(EDDA)(H2O)2] · H2O,

S. Rehman; M. Arshad; K. Masud; R. Afzal; U. Salma

2010-01-01

126

Quantitative analysis of ethynodiol diacetate and ethinyl estradiol/mestranol in oral contraceptive tablets by high-performance liquid chromatography.  

PubMed

A procedure is described for the assay of ethynodiol diacetate and ethinyl estradiol/mestranol by HPLC using two UV detectors at 210 and 280 nm. The system was acetonitrile 38% (v/v) in water as mobile phase on a 250 x 3.2-mm i.d. RP-2 column, with butylated hydroxytoluene as the internal standard. There was greater than 99% recovery from synthetic preparations and the coefficient of variation was greater than 2.0% for formulations. PMID:7062257

Carignan, G; Lodge, B A; Skakum, W

1982-02-01

127

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

EPA Science Inventory

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

128

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.

129

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

130

Mineral Stains at the No Name Prospect  

USGS Multimedia Gallery

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

131

Original article Blue-stain fungi associated  

E-print Network

Original article Blue-stain fungi associated with Tomicus piniperda in Sweden and preliminary to determine the development of blue-staining of sapwood. Fungi were isolated from samples of inner bark isolated from trees attacked by T piniperda. Three species of fungi were rather frequent- ly isolated

Paris-Sud XI, Université de

132

Pro-Q Diamond Phosphoprotein Gel Stain  

E-print Network

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

Lebendiker, Mario

133

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

PubMed Central

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

De Carlo, Sacha; Harris, J. Robin

2010-01-01

134

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

PubMed Central

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

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

2014-01-01

135

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

136

Ultra-widefield fluorescein angiography reveals retinal phlebitis in Susac's syndrome.  

PubMed

A 23-year-old woman with history of headaches and auditory changes presented with acute-onset visual field loss in the right eye. The combination of multiple retinal branch artery occlusions of the right eye on funduscopic examination, characteristic white matter lesions in the corpus callosum on magnetic resonance imaging, and hearing loss on audiometric testing led to a diagnosis of Susac's syndrome. Ultra-widefield fluorescein angiography revealed involvement of the retinal veins, which has not been previously reported with this condition. Additionally, ultra-widefield indocyanine green angiography demonstrated changes in the choroidal circulation, which are controversial in this syndrome. PMID:24972181

Klufas, Michael A; Dinkin, Marc J; Bhaleeya, Swetangi D; Chapman, Kristin O; Riley, Claire S; Kiss, Szilárd

2014-01-01

137

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

138

Purified azure B as a reticulocyte stain.  

PubMed Central

A comparison has been made between reticulocyte preparations stained with purified azure B and with several commerically available batches of brilliant cresyl blue and new methylene blue. Marked variations were observed in the composition and staining performances of the various batches of the two commerically available dyes. Although there were no significant differences in reticulocyte counts obtained with these two dyes, varying amounts of an extraneous, particulate dye deposit were present in these preparations, making accuracte counting both tedious and timeconsuming. Purified azure B, on the other hand, gave reproducibly stained, deposit-free preparations. Reticulocyte counts obtained from azure B preparations correlated almost exactly with those determined using new methylene blue. Purified azure B is therefore recommended as a convenient reticulocyte stain for routine use. Images PMID:64475

Marshall, P N; Bentley, S A; Lewis, S M

1976-01-01

139

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

140

Gram staining apparatus for space station applications.  

PubMed Central

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

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

1990-01-01

141

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

Technology Transfer Automated Retrieval System (TEKTRAN)

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

142

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

Technology Transfer Automated Retrieval System (TEKTRAN)

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

143

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

Technology Transfer Automated Retrieval System (TEKTRAN)

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

144

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

Technology Transfer Automated Retrieval System (TEKTRAN)

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

145

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

Technology Transfer Automated Retrieval System (TEKTRAN)

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

146

Synthesis and characterization of Pt(IV) fluorescein conjugates to investigate Pt(IV) intracellular transformations.  

PubMed

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

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

2013-10-16

147

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, Fite’s acid fast, Giemsa, and H&E stains on eight aquatic microsporidian organisms that were readily available in our two laboratories: Loma salmonae, Glugea anomala, Pseudoloma neurophilia, Pleistophora hyphessobryconis, Pleistophora vermiformis, Glugea sp., Steinhausia mytilovum and an unidentified microsporidian from E. sinensis, UK. Based on tinctorial properties and background staining, the Luna stain performed better for detection of 6 of the 8 microsporidia. Gram stain was superior for the two microsporidia from invertebrates, Steinhausia mytilovum and the unidentified microsporidian from E. sinensis. PMID:21848126

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

2014-01-01

148

A Chemical Stain for Identifying Arsenic-Treated Wood  

E-print Network

A Chemical Stain for Identifying Arsenic-Treated Wood (FINAL) Submitted June 23, 2006 Amy Omae-TREATED WOOD II.1 Applying Phosphate Stains to Arsenate Stains 7 II.2 A Potential Arsenic-Test Kit 14 II.3 Whole Wood Application of the Modified Stannous Chloride Stain 19 II.4 Other Attempted Stain

Florida, University of

149

The influence of staining procedure on differential round cell analysis in stained smears of human semen.  

PubMed

Giemsa and Bryan-Leishman smear staining techniques have been quantitatively evaluated for their ability to determine round cells in human semen. Samples of fertile and vasectomy ejaculates were compared against counts obtained from semithin Araldite sections stained with toluidine blue. TEM studies and immunogold staining of the Pradite section permitted identification and quantitation of nucleated cell profiles. Differential counts from each of the three stains on the same set of semen samples were compared using regression analysis. Counts of seminiferous tubule elements from stain to stain correlated well (r > 0.9). Numerical analyses indicated, however, that leucocytes were commonly misidentified. The r values for neutrophils were less than 0.8 and as low as 0.55 for lymphocytes. These low correlations presumably were due to failures to distinguish between these cells and seminiferous tubule elements. PMID:8724436

Williams, M A; Wick, A; Smith, D C

1996-05-01

150

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

151

The ocular ischemic syndrome. Clinical, fluorescein angiographic and carotid angiographic features.  

PubMed

The records of 43 consecutive patients (51 eyes) with the ocular ischemic syndrome (ocular symptoms and signs attributable to severe carotid artery obstruction) were studied in a retrospective fashion. Men comprised 67% of the group and the mean age at presentation was 64.5 years. In the anterior segment, neovascularization of the iris was observed in 66% of eyes and iritis was noted in 18%. Posterior segment signs included narrowed retinal arteries and dilated, but not tortuous, retinal veins. Mid-peripheral retinal hemorrhages were seen in 80% of eyes, posterior segment neovascularization was observed in 37%, and a cherry red spot was noted in 12%. Fluorescein angiography commonly revealed delayed choroidal and retinal filling, while electroretinography generally demonstrated a reduction in the amplitude of both the a- and b-waves. PMID:3182177

Brown, G C; Magargal, L E

1988-02-01

152

A method for assessment of blood volume parameters in pregnant sheep using fluorescein-labelled dextran.  

PubMed

The assessment of blood volume parameters in clinical and research settings has been limited by methods that involve radioactivity, complex assays or are unreliable. We aimed to design a method for measuring blood volume parameters that was non-radioactive, simple, cheap and reliable. We have used a commercially available fluorescein-labelled 250kDa dextran, a large inert molecule, and have measured dilution of this through the intravascular space of pregnant ewes. From this estimation of plasma volume and measured hematocrit, we have calculated blood volume and red cell volume. The blood volume results are 6% lower than those obtained using radiolabelled red cells, but there is no significant difference in red cell volume between methods. The coefficient of variation for repeated measurements of plasma volume measurements is 3.8%. This is a simple, reliable, cheap and non-radioactive method for estimating blood volume parameters in pregnant sheep, and may prove useful in other settings. PMID:17953987

Rumball, C W H; Van Zijl, P; Rutland, M D; Bloomfield, F H; Harding, J E

2008-01-01

153

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

NASA Astrophysics Data System (ADS)

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

Berger, Jeffrey W.

1998-06-01

154

Detection Of Concrete Deterioration By Staining  

DOEpatents

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

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

1999-09-21

155

Laser treatment of port-wine stains  

PubMed Central

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

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

2015-01-01

156

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

157

Comparison of sulfur hexafluoride, fluorescein and rhodamine dyes and the bacteriophage PRD1 in tracing subsurface flow  

Microsoft Academic Search

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 (SF6), 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 SF6 and PRD-1.

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

2003-01-01

158

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

E-print Network

the first large-scale comparison of a fluorescent dye fluorescein C20H10O5Na2 and a gas sulfur hexafluoride spatial scale m to km , and short time scale hours to days estuarine processes, the inert gas sulfur hexafluoride SF6 is more appropriate for use in large spatial scale tens of km and longer time scale weeks

Ho, David

159

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

ERIC Educational Resources Information Center

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

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

2011-01-01

160

Method for copper staining of germanium crystals  

NASA Technical Reports Server (NTRS)

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

Rivet, E. J.

1969-01-01

161

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

162

The Language of Stained-Glass Windows  

ERIC Educational Resources Information Center

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

Brew, Charl Anne

2010-01-01

163

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

164

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

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

2013-01-01

165

Silver Staining of 2D Electrophoresis Gels Thierry Rabilloud  

E-print Network

and 10 times higher than colloidal Coomassie Blue. However, the first silver staining protocols wereSilver Staining of 2D Electrophoresis Gels Thierry Rabilloud CEA-DSV-iRTSV/LCBM and UMR CNRS Head: Silver staining #12;i. Abstract Silver staining is used to detect proteins after electrophoretic

Paris-Sud XI, Université de

166

Induction of free radicals and tumors in the kidneys of Wistar rats by ferric ethylenediamine-N,N'-diacetate.  

PubMed

An iron chelate, ferric ethylenediamine-N,N'-diacetate [Fe(III)-EDDA], was found to produce hydroxyl radicals with hydrogen peroxide, as determined by both a deoxyribose degradation test and electron spin resonance. Hydroxyl radical production was inhibited not only by adding hydroxyl radical scavengers and catalase, but also by adding superoxide dismutase to the reaction mixture, suggesting that superoxide anion may be involved in the hydroxyl radical production. A single injection of Fe(III)-EDDA (10 mg Fe/kg body wt) to Wistar rats induced thiobarbituric acid reactivity in the kidneys and liver. Repeated injections of Fe(III)-EDDA (10 mg Fe/kg body wt, twice weekly for 3 months) induced a 40% incidence of renal tumors, including renal adenocarcinoma and renal adenoma, 1 year later. These results suggest that Fe(III)-EDDA is an effective free radical producer in vitro and in vivo and that it may be useful in preparing animal models related to iron-dependent free radical damage. The results support our hypothesis that endogenous or exogenous iron, complexed with certain kinds of chelators, promotes free radical-dependent tissue damage and ultimately leads to carcinogenesis in the affected tissue. PMID:8001240

Liu, M; Okada, S

1994-12-01

167

Novel methylene modified cyclohexyl ethylenediamine-N,N'-diacetate ligands and their platinum(IV) complexes. Influence on biological activity.  

PubMed

This paper focuses on the synthesis, characterization and biological activity of new N,N'-methylene modified cyclohexyl ethylenediamine-N,N'-diacetate (edda)-type ligands and their Pt(IV) complexes. Both the ligands and complexes were characterized by infrared, UV-vis, ESI-MS, 1D ((1)H, (13)C, (195)Pt) and 2D (COSY, HSQC, HMBC) NMR spectroscopy and elemental analysis. The possible correlation between the reduction potentials and the cytotoxicity of the complexes was examined. The potential antitumoral activity of all compounds was tested in vitro on human melanoma A375, human glioblastoma U251, human prostate cancer PC3, human colon cancer HCT116, mouse melanoma B16 and mouse colon cancer CT26CL25 cells, as well as primary fibroblasts and keratinocytes. The results obtained revealed strong antitumor potential of the newly synthesized drugs with preserved efficacy against cisplatin resistant lines and less toxicity towards nonmalignant counterparts. The mechanism found to be responsible for the observed tumoricidal action of each synthesized compound was induction of apoptosis generally accompanied with caspase activation. Taken together, the effective response to the treatment of a wide range of different cell lines, including cisplatin resistant subclones, as well as induction of apoptosis, as the mechanism suggested to be the most desirable way of eliminating malignant cells, represents a great advantage of this novel group of drugs in comparison to other members in this metallo-drug family. PMID:22369771

Mihajlovi?, Ljiljana E; Savi?, Aleksandar; Poljarevi?, Jelena; Vu?kovi?, Ivan; Moji?, Marija; Bulatovi?, Mirna; Maksimovi?-Ivani?, Danijela; Mijatovi?, Sanja; Kalu?erovi?, Goran N; Stoši?-Gruji?i?, Stanislava; Miljkovi?, ?or?e; Grguri?-Šipka, Sanja; Sabo, Tibor J

2012-04-01

168

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

169

Improved Whole-Blood-Staining Device  

NASA Technical Reports Server (NTRS)

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

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

2012-01-01

170

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

PubMed Central

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

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

2011-01-01

171

A simple technique for staining of platyhelminths with the lactophnol cotton blue stain.  

PubMed

This paper describes a simple technique for staining of flatworms using lactophenol cotton blue (LPCB). The staining was tested on 2 trematode species: Heterophyes heterophyes and Mesostephanus appendiculatus, and one cestode: Diplopylidium acanthotetra, which were collected from the intestine of stray cats in Kuwait. The specimens were mounted in a small amount of the LPCB stain on a clean slide for 2-3 minutes before covering with a cover slip. The technique rapidly and clearly differentiated the internal structures of the helminthes. Its speed and simplicity are advantages over other staining methods. It is easily used in wide-scale surveys where a large number of platyhelminths have to be identified and it is suitable for field studies. PMID:24260820

Henedi, Adawia A M; El-Azazy, Osama M E

2013-08-01

172

Laser Treatment of Port Wine Stains  

NASA Astrophysics Data System (ADS)

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

Majaron, Boris; Nelson, J. Stuart

173

Lead exposure in stained glass workers  

Microsoft Academic Search

To evaluate lead exposure in stained glass workers, we measured blood lead levels in 12 professional glass workers, in 5 hobbyists, and in 4 workers' family members. Professional workers lead levels (mean 20.7 micrograms\\/dl) were higher than hobbyists' (11.6 micrograms\\/dl) (P . 0.02) or family members' (11.3 micrograms\\/dl). Levels increased with years worked, hours worked per week, and percentage of

Philip J. Landrigan; Peter B. Tamblyn; Mark Nelson; Peter Kerndt; Kenneth J. Kronoveter; Matthew M. Zack

1980-01-01

174

Time-Saving Benefits of Intravital Staining.  

PubMed

One of the challenges in labeling tissues for fluorescence microscopy is minimizing sample processing while maintaining or improving the information generated by the fluorescent label. Generally, tissues are extracted, fixed, and embedded in mounting media (such as paraffin), sectioned, and then postprocessed by removing the paraffin, blocking, labeling, and washing. Despite all of these steps, the consistency of labeling quality can vary as a result of several factors, including heterogeneity in labeling efficiency from slide to slide, the necessity of postprocessing to obtain information on sequential sections of tissue, interference from the mounting media, and loss of native three-dimensional structural information, especially in thicker sections. A method for embedding and processing tissues that have been labeled by intravital staining is described in this study. Intravital staining is the process in which live-cell dyes and other labels are injected into the bloodstream before fixation of the tissues. Tissues processed this way can be imaged upon sectioning without further staining and retain their native, three-dimensional information, thereby improving the information retained by the labels and speeding up sample processing. PMID:20622939

Macgillivray, Catherine; Sylvan, Jeremy; Lee, Richard T; Huang, Hayden

2008-09-01

175

A novel dual-flow bioreactor simulates increased fluorescein permeability in epithelial tissue barriers.  

PubMed

Permeability studies across epithelial barriers are of primary importance in drug delivery as well as in toxicology. However, traditional in vitro models do not adequately mimic the dynamic environment of physiological barriers. Here, we describe a novel two-chamber modular bioreactor for dynamic in vitro studies of epithelial cells. The fluid dynamic environment of the bioreactor was characterized using computational fluid dynamic models and measurements of pressure gradients for different combinations of flow rates in the apical and basal chambers. Cell culture experiments were then performed with fully differentiated Caco-2 cells as a model of the intestinal epithelium, comparing the effect of media flow applied in the bioreactor with traditional static transwells. The flow increases barrier integrity and tight junction expression of Caco-2 cells with respect to the static controls. Fluorescein permeability increased threefold in the dynamic system, indicating that the stimulus induced by flow increases transport across the barrier, closely mimicking the in vivo situation. The results are of interest for studying the influence of mechanical stimuli on cells, and underline the importance of developing more physiologically relevant in vitro tissue models. The bioreactor can be used to study drug delivery, chemical, or nanomaterial toxicity and to engineer barrier tissues. PMID:24756869

Giusti, Serena; Sbrana, Tommaso; La Marca, Margherita; Di Patria, Valentina; Martinucci, Valentina; Tirella, Annalisa; Domenici, Claudio; Ahluwalia, Arti

2014-09-01

176

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

PubMed Central

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

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

2014-01-01

177

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

178

Comparison of the Photobleaching and Photostability Traits of Alexa Fluor 568- and Fluorescein Isothiocyanate- conjugated Antibody  

PubMed Central

Objective: Synthetic fluorescent dyes that are conjugated to antibodies are useful tools to probe molecules. Based on dye chemical structures, their photobleaching and photostability indices are quite diverse. It is generally believed that among different fluorescent dyes, Alexa Fluor family has greater photostability than traditional dyes like fluorescein isothiocyanate (FITC) and Cy5. Alexa Fluor 568 is a member of Alexa Fluor family presumed to have superior photostability and photobleahing profiles than FITC. Materials and Methods: In this experimental study, we conjugated Alexa Fluor 568 and FITC dyes to a mouse anti-human nestin monoclonal antibody (ANM) to acquire their photobleaching profiles and photostability indices. Then, the fluorophore/antibody ratios were calculated using a spectrophotometer. The photobleaching profiles and photostability indices of conjugated antibodies were subsequently studied by immunocytochemistry (ICC). Samples were continuously illuminated and digital images acquired under a fluorescent microscope. Data were processed by ImageJ software. Results: Alexa Fluor 568 has a brighter fluorescence and higher photostability than FITC. Conclusion: Alexa Fluor 568 is a capable dye to use in photostaining techniques and it has a longer photostability when compared to FITC. PMID:23508937

Mahmoudian, Jafar; Hadavi, Reza; Jeddi-Tehrani, Mahmood; Mahmoudi, Ahmad Reza; Bayat, Ali Ahmad; Shaban, Elham; Vafakhah, Mohtaram; Darzi, Maryam; Tarahomi, Majid; Ghods, Roya

2011-01-01

179

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

180

Dendritic cells and the initiation of contact sensitivity to fluorescein isothiocyanate.  

PubMed Central

Lymph node cells taken 24 hr after skin-painting mice with the contact sensitizer fluorescein isothiocyanate (FITC) induce delayed-type hypersensitivity in recipient mice. Skin-painting increased the number of dendritic cells (DC) in the draining lymph nodes without significantly changing the number of lymphocytes at 24 hr. The antigen was preferentially located on the DC. Raising the dose of FITC increased both the number of DC and the amount per cell. The addition of these DC to syngeneic lymph node cells at a ratio as low as 1:300 initiated proliferative responses in vitro. The level of proliferation was related to the amount of antigen on the DC. Mice given 50,000 of these fluorescent DC developed specific contact sensitivity reactions. DC exposed in vitro to FITC also acquired antigen and were able to initiate proliferative responses in vitro and to sensitize recipient mice. The DC may therefore be the prime cell involved in the induction of delayed-type hypersensitivity. PMID:3100437

Macatonia, S E; Edwards, A J; Knight, S C

1986-01-01

181

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

182

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

PubMed Central

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

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

2002-01-01

183

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

PubMed

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

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

2014-07-01

184

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

185

5-(Pentafluorobenzoylamino)fluorescein: A Selective Substrate for the Determination of Glutathione Concentration and Glutathione S-Transferase Activity  

Microsoft Academic Search

5-(Pentafluorobenzoylamino)fluorescein (PFB-F), a new thiol-reactive molecule was synthesized to improve the detection limits and specificity of the assays for glutathioneS-transferase (GST) activity and glutathione (GSH). A rapid assay method to measure GSH concentration or GST activity and the simultaneous analysis of multiple samples is possible because the glutathione adduct, GS-TFB-F, is separated from PFB-F by thin-layer chromatography (TLC) and can

Seksiri Arttamangkul; Mahesh K. Bhalgat; Rosaria P. Haugland; Zhenjun Diwu; Jixiang Liu; Dieter H. Klaubert; Richard P. Haugland

1999-01-01

186

Use of stains to detect fingermarks.  

PubMed

Detection of fingermarks at a crime scene or on related items is of prime interest for forensic investigators, mainly for identification purposes. Most of the fingermarks are invisible to the naked eye, however. The application of detection techniques is required to establish visual contrast between the secretion residue and the underlying substrate. We give here a review of the field related to the concept of using stains to detect fingermarks. A distinction has been made between the physically driven classical detection techniques, the chemically driven ones, and those based on nanostructured materials, an emerging field in forensic science. PMID:20121464

Becue, A; Moret, S; Champod, C; Margot, P

2011-06-01

187

Hyphal walls of isolated lichen fungi: autoradiographic localization of precursor incorporation and binding of fluorescein-conjugated lectins.  

PubMed

The hyphal walls of three mycobionts, isolated from the lichens Xanthoria parietina, Tornabenia intricata and Sarcogyne sp. were investigated by two techniques: microautoradiography of fungal colonies exposed to radioactive carbohydrate precursors: and binding, in vivo, of fluorescein conjugated lectins to hyphal walls of such colonies. N-[3H] acetylglucosamine was readily incorporated into tips, young hyphal walls and septa of the three mycobionts and the free-living fungus Trichoderma viride, but not into Phytophthora citrophthora, indicating that chitin is a major component of the mycobionts' hyphal walls. All three mycobionts, but neither of the free-living fungi, incorporated [3H] mannose and [3H] mannitol into their hyphal walls. Fluorescein-conjugated wheat germ agglutinin was bound to the hyphal walls of the three mycobionts and T. viride, but not to the walls of P. citrophthora; the binding pattern was similar to the grain pattern obtained in autoradiographs after short N-[3H]acetylglucosamine labelling. As wheat germ agglutinin binds specifically to chitin oligomers, the lectin binding tests further confirmed that chitin is a mycobiont hyphal wall component. Binding characteristics of several fluorescein-conjugated lectins to the three mycobionts indicated that this technique can yield useful information concerning the chemical composition of hyphal wall surfaces. PMID:1275648

Galun, M; Braun, A; Frensdorff, A; Galun, E

1976-05-01

188

Influence of topical cyclosporine A and dissolvent on corneal epithelium permeability of fluorescein.  

PubMed

The corneal stroma is the major barrier to penetration for the lipophilic Cyclosporine A (CsA) molecule and prevents the use of the common ophthalmic solvents. At present, corn oil, castor oil and olive oil are the three most commonly used vehicles. The aim of this study was to determine the effect that topically applied CsA dissolved in different oils has on corneal epithelial permeability measured by fluorophotometry. Forty healthy volunteers, with absence of ocular or systemic disease and not receiving topical or systemic drugs were enrolled. Measurements were taken before and 45 min after the instillation of 40 microliters of a 2% aqueous solution of sodium fluorescein without preservatives. Basal corneal permeability and the permeability 24 h after the instillation of 2% CsA-olive oil, olive oil alone, 2% CsA-castor oil, castor oil alone, 2% CsA-corn oil and corn oil alone, were calculated. To prepare the topical 2% CsA, a Sandimmun oral solution (Sandoz, Basel, Switzerland) was employed under sterile conditions. We found that epithelial permeability 24 h after the instillation of any CsA formulations or solvents increased more than 6.62 times (p <0.001). No differences in corneal permeability values were found between any of the CsA formulations and the vehicles. We conclude that oils used to dissolve CsA are mainly responsible for the increased corneal epithelial permeability. No differences were found in the effects of the tested solvents on corneal epithelial permeability. PMID:8861636

Benítez del Castillo, J M; Castillo, A; Toledano, N; Durán, S; del Aguila, C; Otero, M; García-Sanchez, J

1995-01-01

189

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2011 CFR

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

2011-04-01

190

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2012 CFR

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

2012-04-01

191

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2013 CFR

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

2013-04-01

192

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2014 CFR

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

2014-04-01

193

Treatment of port-wine stains: analysis.  

PubMed

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 CO2 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. PMID:3452741

van Gemert, M J; Welch, A J

1987-08-01

194

Detection of Mycobacterial Infections Using the Dieterle Stain  

Microsoft Academic Search

Retrospective review comparing the modified Dieterle stain with standard acid-fast stains was performed on seven surgical\\u000a pathology cases that contained culture-positive mycobacteria infections. Tissues examined comprised cervical and submandibular\\u000a lymph nodes and soft tissues of the face and chest. Modified Dieterle staining was performed on paraffin-embedded tissue sections,\\u000a and the results were compared with those of hematoxylin-eosin stains and auramine-rhodamines

John G. Brady; Gordon E. Schutze; Robert Seibert; Hazel V. Horn; Barbara Marks; David M. Parham

1998-01-01

195

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

NASA Astrophysics Data System (ADS)

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

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

2015-02-01

196

Machine Vision System for Automated Detection of Stained Pistachio Nuts  

Microsoft Academic Search

A machine vision system was developed to separate stained pistachio nuts, which comprise about 5% of the California crop, from unstained nuts. Stained nuts have lower consumer acceptance and higher incidences of aflatoxin contamination. The machine vision system may be used as an automated quality control device to detect stained nuts. The system was tested on three different pistachio process

Tom Pearson

1996-01-01

197

bleachingIn vitro chemical stain removal by 'whitening' toothpastes  

Microsoft Academic Search

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

Diana Scarrott

2000-01-01

198

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

PubMed Central

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

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

2011-01-01

199

Efficacy of rapid, economical, acetic acid, Papanicolaou stain in cervical smears as an alternative to conventional Papanicolaou stain  

PubMed Central

Background: Papanicolaou (Pap) stain has been used over the years for cervical cytology screening. However; it utilizes a considerable amount of alcohol which is expensive and difficult to procure. In one of the modifications, ethyl alcohol is replaced by 1% acetic acid and is termed as rapid, economical, acetic acid Papanicolaou (REAP) stain. It is cost effective, easily available and provides a suitable and rapid staining alternative. Aim: This study was undertaken to assess the efficacy of REAP stain as an alternative method to conventional Pap stain. Materials and Methods: This study was done over a period of 18 months in a tertiary care hospital. Two sets of cervical smears were prepared of which one was stained with conventional Pap stain, and other was stained with REAP stain. The smears were examined for cytomorphological parameters and were evaluated using a modification of parameters given by Ng et al. Results: A total of 737 smears were examined in duplicate. Most of the conventional Pap smears showed excellent preservation (91.6%) with very few showing optimal (7.6%) and sub-optimal staining (0.8%). In contrast to this excellent preservation was seen in just 33.6% of the REAP stained smears with majority showing optimal and sub-optimal preservation (46.5% and 20% respectively). The P value was statistically significant (<0.0001) depicting inferior staining quality of REAP stain. Conclusion: Rapid, economical, acetic acid Papanicolaou stain undoubtly is a simple, fast and cost effective stain which can be adopted mainly in resource limited settings, but cannot be utilized for research purpose in a tertiary care setup due to poor preservation of the staining quality.

Izhar, Shabnam; Kaur, Rupinder; Masih, Kanwal

2014-01-01

200

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

201

Automated single-slide staining device. [in clinical bacteriology  

NASA Technical Reports Server (NTRS)

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

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

1975-01-01

202

Scrub typhus hepatitis confirmed by immunohistochemical staining  

PubMed Central

Scrub typhus is an acute febrile disease caused by Orientia tsutsugamushi (O. tsutsugamushi). We report herein the case of a woman who presented with fever and elevated serum levels of liver enzymes and who was definitively diagnosed with scrub typhus by histopathological examination of liver biopsy specimens, serological tests and nested polymerase chain reaction. Immunohistochemical staining using a monoclonal anti-O. tsutsugamushi antibody showed focally scattered positive immunoreactions in the cytoplasm of some hepatocytes. This case suggests that scrub typhus hepatitis causes mild focal inflammation due to direct liver damage without causing piecemeal necrosis or interface hepatitis. Thus, scrub typhus hepatitis differs from acute viral hepatitis secondary to liver damage due to host immune responses, which causes severe lobular disarray with diffuse hepatocytic degeneration, necrosis and apoptosis as well as findings indicative of hepatic cholestasis, such as hepatic bile plugs or brown pigmentation of hepatocytes. PMID:23049227

Chung, Jong-Hoon; Lim, Sung-Chul; Yun, Na-Ra; Shin, Sung-Heui; Kim, Choon-Mee; Kim, Dong-Min

2012-01-01

203

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

204

Hydration of an Oriental Spruce (Picea orientalis) Pollen Grain  

NSDL National Science Digital Library

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

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

2004-03-09

205

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

206

Sensitivity of biphotonic systems to light intensity fluctuations: Experimental evidence in the thermoluminescence of fluoresceine in boric acid glass  

SciTech Connect

We study the response of biphotonic systems to fluctuations of the incident light intensity. A convenient experimental model for such a system is the thermoluminescence of fluoresceine in boric acid glass. The location of the steady states of this system as a function of the incident light intensity is determined for constant and fluctuating modes of illumination. A comparison of the results for these two modes reveals that the light intensity fluctuations can significantly modify the curve of the steady states. For low values of the average intensity the efficiency of the photoionization is lowered, whereas for high values of the average light intensity the efficiency is enhanced.

Micheau, J.C.; Horsthemke, W.; Lefever, R.

1984-09-01

207

Synthesis and Purification of a Hammerhead Ribozyme and a Fluorescein-Labeled RNA Substrate. A Biochemistry Laboratory: Part 1  

NASA Astrophysics Data System (ADS)

The applications of in vitro transcription and chemical synthesis of RNA are discussed. This laboratory describes the in vitro synthesis of a 38-nucleotide hammerhead ribozyme and the synthesis of a 17-nucleotide fluorescein-labeled RNA substrate by using standard phosphoramidite methodologies, two widely used methods in modern RNA research. The synthesis and purification procedures outlined allow students to develop an understanding of RNA handling procedures, synthesis of modified nucleic acids, gel electrophoresis, visualization of RNA by nonradioactive techniques, and quantitation of nucleic acids. The RNAs that are synthesized have applications in biotechnology and medicine; thus the students gain access to current problems in chemical and clinical research.

Chow, Christine S.; Somne, Smita

1999-05-01

208

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

PubMed Central

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

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

2011-01-01

209

Fluorescein-labeled "arch-like" DNA probes for electrochemical detection of DNA on gold nanoparticle-modified gold electrodes.  

PubMed

In the present study, a gold nanoparticle-modified gold electrode (nanogold electrode) was used to develop a novel fluorescein electrochemical DNA biosensor based on a target-induced conformational change. The nanogold electrode was obtained by electrodepositing gold nanoparticles onto a bare gold electrode. This modification not only immobilized probe oligonucleotides, but also adsorbed fluorescein onto the surface of the gold nanoparticles to form an "arch-like" structure. This article compares the electrochemical signal changes caused by the hybridization of "arch-like" DNA on nanogold electrode and linear DNA on bare gold electrode. The results showed that the adsorption effect of nanogold can enhance the sensitivity of the sensor. The linear range of target ssDNA is from 2.0 × 10(-9)M to 2.0 × 10(-8)M with a correlation coefficient of 0.9956 and detection limit (3?) of 7.10 × 10(-10)M. Additionally, the specificity and hybridization response of this simple sensor were investigated. PMID:24140637

Xu, Lin; Su, Hao-ra; Sun, Gui-Rong; Wang, Yan; Guo, Shang-jing; Zhang, Xiao-ru; Zhang, Shu-sheng; Xing, Shi-chao

2013-12-01

210

Extrinsic stain removal with a toothpowder: A randomized controlled trial  

PubMed Central

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

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

2014-01-01

211

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

PubMed

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

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

2011-08-01

212

7 CFR 3201.87 - Wood and concrete stains.  

Code of Federal Regulations, 2014 CFR

...2014-01-01 2014-01-01 false Wood and concrete stains. 3201.87 Section...PROCUREMENT Designated Items § 3201.87 Wood and concrete stains. (a) Definition...be applied as a finish for concrete and wood surfaces and that contain dyes or...

2014-01-01

213

7 CFR 3201.87 - Wood and concrete stains.  

Code of Federal Regulations, 2013 CFR

...2013-01-01 2013-01-01 false Wood and concrete stains. 3201.87 Section...PROCUREMENT Designated Items § 3201.87 Wood and concrete stains. (a) Definition...be applied as a finish for concrete and wood surfaces and that contain dyes or...

2013-01-01

214

Particle motion and stain removal during simulated abrasive tooth cleaning  

Microsoft Academic Search

Stain removal from teeth is important both to prevent decay and for appearance. This is usually achieved using a filament-based toothbrush with a toothpaste consisting of abrasive particles in a carrier fluid. This work has been carried out to examine how these abrasive particles interact with the filaments and cause material removal from a stain layer on the surface of

R. Lewis; S. C. Barber; R. S. Dwyer-Joyce

2007-01-01

215

Negative Stains Containing Trehalose: Application to Tubular and Filamentous Structures  

NASA Astrophysics Data System (ADS)

Several examples are presented that show the successful application of uranyl acetate and ammonium molybdate negative staining in the presence of trehalose for TEM studies of filamentous and tubular structures. The principal benefit to be gained from the inclusion of trehalose stems from the considerably reduced flattening of the large tubular structures and the greater orientational freedom of single molecules due to an increased depth of the negative stain in the presence of trehalose. Trehalose is likely to provide considerable protection to protein molecules and their assemblies during the drying of negatively stained specimens. Some reduction in the excessive density imparted by uranyl acetate around large assemblies is also achieved. Nevertheless, in the presence of 1% (w/v) trehalose, it is desirable to increase the concentration of negative stain to 5% (w/v) for ammonium molybdate and to 4% for uranyl acetate to produce satisfactory image contrast. In general, the ammonium molybdate-trehalose negative stain is more satisfactory than the uranyl acetate-trehalose combination, because of the greater electron beam sensitivity of the uranyl negative stain. Reassembled taxol-stabilized pig brain microtubules, together with collagen fibrils, sperm tails, helical filaments, and reassociated hemocyanin (KLH2), all from the giant keyhole limpet Megathura crenulata, have been studied by negative staining in the presence of trehalose. In all cases satisfactory TEM imaging conditions were readily obtained on the specimens, as long as regions of excessively deep stain were avoided.

Harris, J. Robin; Gerber, Max; Gebauer, Wolfgang; Wernicke, Wolfgang; Markl, Jürgen

1996-02-01

216

OBSERVATION OF SALMONELLA TYPHIMURIUM FIMBRIAE BY NEGATIVE STAIN  

Technology Transfer Automated Retrieval System (TEKTRAN)

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

217

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 blue–directed biopsy in detecting specialized columnar epithelium and dysplasia in Barrett's esophagus. We performed upper endoscopy with methylene blue–directed biopsy and obtained 236 large cup biopsy specimens (145 stained,

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

1996-01-01

218

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

219

A new cleaning method for historic stained glass windows  

Microsoft Academic Search

Historical stained glass has a clear tendency to form a crusted layer on its surface due to the environmental exposure. One of the most delicate aspects to be faced during the restoration of historic glass windows is the cleaning of these thick corrosion crusts.For several centuries, stained glass windows have been cleaned using damaging mechanical (scalpel) and chemical (high acidic

Sonia Murcia-Mascarós; Paola Foglia; M. Laura Santarelli; Clodoaldo Roldán; Rafael Ibañez; Alfonso Muñoz; Pablo Muñoz

2008-01-01

220

New series of Tc-99m-labeled hepatobiliary tracers: N'-acyl- and N'-sulfonyl ethylenediamine-N,N-diacetic acids  

SciTech Connect

Various Tc-99m-labeled N'-substituted derivatives of ethylenediamine-N,N-diacetic acid (EDDA) are evaluated as hepatobiliary imaging agents. N-substituted aromatic acyl and aromatic sulfonyl derivatives of EDDA, labeled with Tc-99m, were administered to rabbits and golden hamsters, and the distribution indicated clearance by the hepatobiliary system. N'-aromatic sulfonyl EDDAs were labeled with Tc-99m by the SnCl/sub 2/ method with more than 99% yield. Clearance of Tc-99m-p-toluenesulfonyl EDDA from the blood and the liver was as rapid as that of TC-99m N-(2,6-diethylphenylcarbamoylmethyl)iminodiacetic acid (Tc-99m benzenesulfonyl EDDA lowered urinary excretion. It is concluded that the sulfonyl EDDAs provide a fruitful source for Tc-99m-labeled hepatobiliary radiopharmaceuticals.

Karube, Y.; Kono, A.; Maeda, T.; Ohya, M.; Matsushima, Y.

1981-07-01

221

New series of Tc-99m-labeled hepatobiliary tracers: N'-acyl- and N'-sulfonyl ethylenediamine-N,N-diacetic acids  

SciTech Connect

Various Tc-99m-labeled N'-substituted derivatives of ethylenediamine-N,N-diacetic acid (EDDA) are evaluated as hepatobiliary imaging agents. N'-substituted aromatic acyl and aromatic sulfonyl derivatives of EDDA, labeled with Tc-99m, were administered to rabbits and golden hamsters, and the distribution indicated clearance by the hepatobiliary system. N'-aromatic sulfonyl EDDAs were labeled with Tc-99m by the SnCl/sub 2/ method with more than 99% yield. Clearance of Tc-99m-p-toluenesulfonyl EDDA from the blood and the liver was as rapid as that of Tc-99m N-(2,6-diethylphenylcarbamoylmethyl)iminodiacetic acid (Tc-99m diethyl IDA). Substitution of a bulky group at the aromatic ring in Tc-99m benzene-sulfonyl EDDA lowered urinary excretion. It is concluded that the sulfonyl EDDAs provide a fruitful source for Tc-99m-labeled hepatobiliary radiopharmaceuticals.

Karube, Y.; Kono, A.; Maeda, T.; Ohya, M.; Matsushima, Y.

1981-07-01

222

In vivo photoacoustic imaging of model of port wine stains.  

PubMed

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

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

2012-01-01

223

Mapping stain distribution in pathology slides using whole slide imaging  

PubMed Central

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

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

2014-01-01

224

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

225

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. PMID:24727376

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

2014-01-01

226

Staining with methylthioninium chloride for the diagnosis of fungal keratitis  

PubMed Central

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

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

2013-01-01

227

Staining of intracellular deposits of uranium in cultured murine macrophages.  

PubMed

In our studies of the health effects of internalized depleted uranium, we developed a simple and rapid light microscopic method to stain specifically intracellular uranium deposits. Using J774 cells, a mouse macrophage line, treated with uranyl nitrate and the pyridylazo dye 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol, uranium uptake by the cells was followed. Specificity of the stain for uranium was accomplished by using masking agents to prevent the interaction of the stain with other metals. Prestaining wash consisting of a mixture of sodium citrate and ethylenediaminetetraacetic acid eliminated staining of metals other than uranium. The staining solution consisted of the pyridylazo dye in borate buffer along with a quaternary ammonium salt, ethylhexadecyldimethylammonium bromide, and the aforementioned sodium citrate/ethylenediaminetetraacetic acid mixture. The buffer was essential for maintaining the pH within the optimum range of 8 to 12, and the quaternary ammonium salt prevented precipitation of the dye. Staining was conducted at room temperature and was complete in 30 min. Staining intensity correlated with both uranyl nitrate concentration and incubation time. Our method provides a simple procedure for detecting intracellular uranium deposits in macrophages. PMID:11871745

Kalinich, J F; McClain, D E

2001-01-01

228

C4d staining as immunohistochemical marker in inflammatory myopathies.  

PubMed

The diagnosis of an inflammatory myopathy is often established based on basic histologic studies. Additional immunohistochemical studies are sometimes required to support the diagnosis and the classification of inflammatory myopathies. Staining for major histocompatibility complex 1 (MHC1) often shows increased sarcolemmal labeling in inflammatory myopathies. Endomysial capillary staining C5b-9 (membrane attack complex) is a feature that is reported as frequently associated with dermatomyositis. Immunohistochemical staining for C4d is widely used for various applications including the assessment of antibody-mediated rejection after solid organ transplantation. In the context of dermatomyositis, C4d staining has been described in skin biopsies but not in muscle biopsies. A total of 32 muscle biopsy specimens were examined. The hematoxylin and eosin-stained slides were reviewed, and immunohistochemical studies for MHC1, C5b-9, and C4d were conducted. The staining observed for C5b-9 and C4d was compared. Overall, the staining pattern for C4d mirrored the one observed for C5b-9 in the examined muscle biopsy specimens. There was high and statistically significant (P<0.0001) correlation between the staining seen with these 2 antibodies. Both antibodies labeled the cytoplasm of degenerating necrotic myofibers. In addition, both antibodies showed distinct endomysial capillary labeling in a subset of dermatomyositis. Areas with perifascicular atrophy often exhibited the most prominent vascular labeling for C4d and C5b-9. In conclusion, C4d and C5b-9 show similar expression patterns in muscle biopsies of patients with inflammatory myopathies and both highlight the presence of vascular labeling associated with dermatomyositis. C4d antibodies are widely used and may offer an alternative for C5b-9 staining. PMID:24897075

Pytel, Peter

2014-10-01

229

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

PubMed Central

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

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

2013-01-01

230

Spindle microtubule dynamics in sea urchin embryos: analysis using a fluorescein-labeled tubulin and measurements of fluorescence redistribution after laser photobleaching  

Microsoft Academic Search

The rate of exchange of tubulin that is incorporated into spindle microtubules with dimeric tubulin in the cytoplasm has been measured in sea urchin eggs by studying fluores- cence redistribution after photobleaching (FRAP). Dichlorotriazinyl amino fluorescein (DTAF) has been used to label bovine brain tubulin. DTAF-tubulin has been injected into fertilized eggs of Lytechinus variegatus and allowed to equilibrate with

E. D. Salmon; R. J. LESLIE; W. M. SAXTON; M. L. KAROW; J. R. MclNTOSH

1984-01-01

231

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

Microsoft Academic Search

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

Jill M. Picton; Martin W. Steer

1985-01-01

232

Improving acid-fast fluorescent staining for the detection of mycobacteria using a new nucleic acid staining approach.  

PubMed

Acid fast staining of sputum smears by microscopy remains the prevalent method for detecting Mycobacterium tuberculosis. The sensitivity of microscopy using acid fast stains requires 10(4) bacilli per ml of sputum. Although fluorescent acid fast stains, such as Auramine-O, show improved sensitivity, almost half of culture-positive TB cases are currently estimated to remain smear-negative. These current diagnosis problems provide impetus for improving staining procedures. We evaluated a novel fluorescent acid-fast staining approach using the nucleic acid-binding dye SYBR(®) Gold on mycobacterial in vitro cultures. The SYBR(®) Gold stain detected 99% of MTB in both actively replicating aerobic and non-replicating hypoxic cultures. Transmission light microscopy with Ziehl-Neelsen fuchsin, and fluorescence microscopy with Auramine-O or Auramine-rhodamine detected only 54%-86% of MTB bacilli. SYBR(®) Gold fluoresces more intensely than Auramine-O, and is highly resistant to fading. The signal to noise ratio is exceptionally high due to a >1000-fold enhanced fluorescence after binding to DNA/RNA, thereby reducing most background fluorescence. Although cost and stability of the dye may perhaps limit its clinical use at this time, these results warrant further research into more nucleic acid dye variants. In the meantime, SYBR(®) Gold staining shows great promise for use in numerous research applications. PMID:25130623

Ryan, Gavin J; Shapiro, Howard M; Lenaerts, Anne J

2014-09-01

233

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

234

Immunohistochemical staining for ranaviruses Introduction: Ranaviruses negatively impact amphibian populations  

E-print Network

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

Gray, Matthew

235

A cold staining method for acid-fast bacilli  

PubMed Central

The Ziehl-Neelsen method is probably the best known and most frequently used procedure for staining tubercle bacilli. The method requires controlled heating for its success. However, in developing countries, such as India, where most laboratories rely mainly on spirit lamps as a source of heat, the Ziehl-Neelsen method often cannot be carried out because rectified spirit is difficult to obtain. The study describes a cold staining technique that uses the same staining solutions as the conventional Ziehl-Neelsen method. For direct smears, the correlation of results of the cold staining procedure with those of the Ziehl-Neelsen method was 97% and for concentrated smears was 99%. The method described is suitable for use in basically equipped laboratories. PMID:2433067

Vasanthakumari, R.; Jagannath, K.; Rajasekaran, S.

1986-01-01

236

21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2010 CFR

21 Food and Drugs 8 2010-04-01 2010-04-01... 864.1850 Section 864.1850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT... HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and...

2010-04-01

237

Pyronin Y as a Fluorescent Stain for Paraffin Sections  

Microsoft Academic Search

Pyronin Y has long been used, in combination with other dyes such as Methyl Green, as a differential stain for nucleic acids\\u000a in paraffin tissue sections. It also forms fluorescent complexes with double-stranded nucleic acids, especially RNA, enabling\\u000a semi-quantitative analysis of cellular RNA in flow cytometry. However, the possibility of using pyronin Y as a fluorescent\\u000a stain for paraffin tissue

Bo Li; Ying Wu; Xiao-Ming Gao

2002-01-01

238

Detection and identification of asbestos by microscopical dispersion staining  

PubMed Central

Asbestos fibers as small as 1 ?m in diameter can be uniquely identified by light microscopy by employing dispersion staining methods. The technique described herein involves suspension of fibers in liquids of known refractive indices and observation of color display by means of a dispersion staining objective. Wavelengths or indices of refraction may be determined at right angles to and parallel to fiber axes. This method is rapid and sensitive for identification purposes. Imagesp[61]-a PMID:4470956

McCrone, Walter C.

1974-01-01

239

Application of a vital fluorescent staining method for simultaneous, near-real-time concentration monitoring of two bacterial strains in an Atlantic coastal plain aquifer in Oyster, Virginia.  

PubMed

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

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

2004-03-01

240

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

PubMed Central

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

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

2004-01-01

241

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

E-print Network

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

Mecham, Robert

242

Reliability of a rapid hematology stain for sputum cytology*  

PubMed Central

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

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

2014-01-01

243

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

244

Plants can utilize iron form Fe?N,N'?di?(2?hydroxybenzoyl)?ethylenediamine?N,N'?diacetic acid, a ferric chelate with 10greater formation constant than Fe?EDDHA  

Microsoft Academic Search

HBED [N,N'?di?(2?hydroxybenzoyl)?ethylenediamine?N,N'?diacetic acid] was synthesized in 1967, but it only recently became available in the quantities needed for plant nutrition research. This chelator is a close relative of EDDHA, but has a Fe?chelate formation or stability constant (at ionic strength, ?+ 0.1) of 39.68 compared to 33.91 for EDDHA. Even the Fe selectivity (i.e. Fe\\/Ca stability constant ratios) is greater

Rufus L. Chaney

1988-01-01

245

Amyloid Histology Stain for Rapid Bacterial Endospore Imaging ? †  

PubMed Central

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

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

2011-01-01

246

[Usefulness and limit of Gram staining smear examination].  

PubMed

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

Nagata, Kuniaki; Mino, Hirotoshi; Yoshida, Shunsuke

2010-05-01

247

Coomassie blue staining for high sensitivity gel-based proteomics.  

PubMed

Gel electrophoresis, particularly one- (1DE) and two-dimensional electrophoresis (2DE), remain among the most widely used top-down methods for resolving and analysing proteomes. Detection of the resulting protein maps relies on staining (i.e. colloidal coomassie blue (CCB) or SYPRO Ruby (SR), in addition to many others). Fluorescent in-gel protein stains are generally preferred for higher sensitivity, reduced background, and wider dynamic range. Although traditionally used for densitometry, CBB has fluorescent properties. Indeed, infrared detection of CCB stained protein was comparable to SR, with BioSafe (Bio-Rad) and the Neuhoff formulation (NCCB) identified as potentially superior to SR; a minor sensitivity issue encountered in gel-resolved proteomes; might have been due to the unified staining protocol used. Here the staining protocol for both CCB formulations was optimised, yielding improved selectivity without affecting sensitivity; the resulting linear dynamic range was similar for BioSafe and NCCB and somewhat better than SR. 2D gel-based analyses of mouse brain and Arabidopsis thaliana (leaf) proteomes indicated markedly superior spot detection using the NCCB formulation. Thus more sensitive, quantitative in-gel protein analyses can be achieved using NCCB, at a fraction of the cost. PMID:23428344

Gauci, Victoria J; Padula, Matthew P; Coorssen, Jens R

2013-09-01

248

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. PMID:21286387

Pérez Gomes, Jessica

2011-01-01

249

Specific staining of tissue components with metal-hematoxylin complexes.  

PubMed

The older metal-hematoxylin stains stain a broad spectrum of tissue components. Several recently introduced metal-hematoxylin stains are highly selective. This selectivity is usually bought at the price of severe limitations on the choice of fixative. A very dilute (2 x 10(-4)M) aluminum hematoxylin is selective for nucleic acids in tissues fixed in organic solvents alone. Vanadate hematoxylin is selective for basic proteins in tissues fixed in formaldehyde or mercuric salts. Bismuth hematoxylin is selective for arginine residues and thus for histones and myelin basic protein in tissues fixed in strong acids (Bouin's fluid or SUSA fluid). Zirconyl hematoxylin is selective for acidic mucins. Zirconyl hematoxylin does not restrict the choice of fixative. PMID:11473818

Smith, A A

2002-01-01

250

A staining protocol for identifying secondary compounds in Myrtaceae1  

PubMed Central

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

Retamales, Hernan A.; Scharaschkin, Tanya

2014-01-01

251

Synthesis of adenosine derivatives as transcription initiators and preparation of 5? fluorescein- and biotin-labeled RNA through one-step in vitro transcription  

PubMed Central

Expanding our previous finding of an adenosine-initiated transcription system, we now demonstrate that either the 5? site or the N6 site of adenosine nucleotides can be modified extensively without abolishing their ability to initiate transcription under the T7 ?2.5 promoter. Two series of amino derivatives of adenosine nucleotides were synthesized. Fluorescein and biotin groups were coupled to AMP derivatives through linkers of different sizes and hydrophobicities. Both fluorescein- and biotin-conjugated (at either the 5? or N6 site) adenosine nucleotides can act as efficient transcription initiators, producing fluorescein- and biotin-labeled RNA at the specific 5? end by a one-step transcription procedure, eliminating posttranscriptional modification. Furthermore, N6-modified adenosine derivative-initiated transcription synthesizes 5? end modified RNA with a free phosphate group, providing the possibility for further derivatization. The current finding makes easily available a variety of site-specifically functionalized RNA, which may be used in nucleic acid detection, RNA structural and functional investigation, and generation and isolation of novel functional RNA. PMID:14624011

HUANG, FAQING; WANG, GUOCAN; COLEMAN, TRICIA; LI, NA

2003-01-01

252

Antibody Testing Against Canine Coronavirus by Immunoperoxidase Plaque Staining  

Microsoft Academic Search

The application of the immunoperoxidase (IP) plaque staining procedure (IP test) to the diagnosis of canine coronavirus (CCV) infection was investigated. The IP test did not react with sera from either 15 specific pathogen-free (SPF) dogs or 7 SPF dogs immunized with a multivalent vaccine, including canine parvovirus type 2, canine distemper virus, canine adenovirus type 2, and canine parainfluenza

T. Soma; M. Hara; H. Ishii; S. Yamamoto

2001-01-01

253

Sex, Race, and the Stained-Glass Window  

Microsoft Academic Search

“Sex, Race and the Stained-Glass Window” is one person's reflections of her life as an African-American lesbian growing up in a Christian fundamentalist church. The strong currents of racism, sexism, and homophobia flow from rivers of tradition and patriarchal social structures that make one either a strong opponent of those forces or a proponent of the status quo. The author

Nzinga Shaka Zulu

1996-01-01

254

Staining potential of sealants in/on exterior wall substrates  

SciTech Connect

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

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

1996-12-31

255

Rapid enumeration of respiratory active mycobacteria with fluorescent double staining.  

PubMed

A new method for rapid enumeration of physiologically active mycobacteria was developed by acid-fast bacilli staining (Auramine O) following formazan reduction. Results can be obtained within 90 min by the optimized procedure, while more than one week is required for the widely used culture-dependent approach. PMID:20600362

Ichijo, Tomoaki; Izumi, Yoko; Yamaguchi, Nobuyasu; Nasu, Masao

2010-09-01

256

Banding patterns in newt chromosomes by the giemsa stain  

Microsoft Academic Search

Specific banding patterns can be produced on the mitotic chromosomes of the newt species Triturus vulgaris meridionalis and T. italicus by using the Giemsa stain technique. These bands are most useful cytogenetic markers in karyotyping, since they facilitate identification of the individual elements of the complements. Evaluation of the shape of chromosomes as well as of the banding patterns produced

Irma Nardi; Matilde Ragghianti; Giorgio Mancino

1973-01-01

257

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

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

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

258

ENHANCED DAPI STAINING FOR CRYPTOSPORIDIUM IN WATER SAMPLES  

EPA Science Inventory

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

259

A Genetic Algorithm for Grammars James Anderson and Joe Staines  

E-print Network

A Genetic Algorithm for Grammars James Anderson and Joe Staines July 1, 2010 Background training data. 1 #12;A Genetic Algorithm for Grammars Of course, there are many more grammars than be able to search heuristically. Project Proposal We propose a project which uses a genetic algorithm

Goldschmidt, Christina

260

STAINING OF TISSUE SECTIONS FOR ELECTRON MICROSCOPY WITH HEAVY METALS  

Microsoft Academic Search

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

M. L. Watson

1958-01-01

261

MP 33354 Pro-Q Sapphire 532 Oligohistidine Gel Stain  

E-print Network

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

Lebendiker, Mario

262

Analysis of human platelet glycoprotein IIb-IIIa by fluorescein isothiocyanate-conjugated disintegrins with flow cytometry.  

PubMed

Disintegrins are a group of snake venom peptides which inhibit human platelet aggregation by acting as glycoprotein IIb-IIIa (GPIIb-IIIa) antagonists. They are cysteine-rich, Arg-Gly-Asp (RGD)-containing peptides, and bind to GPIIb-IIIa complex on platelet membrane with a very high affinity (Kd, 10(-7)-10(-8) M). In this study, we analyzed GPIIb-IIIa complex on platelet membrane by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated disintegrins as probes. Of these FITC-conjugated disintegrins, FITC-Rhodostomin is the most sensitive probe because Rhodostomin was conjugated with more FITC molecules than Trigramin and Halysin were. The binding fluorescence intensity of FITC-Trigramin (FITC-Tg), FITC-Halysin (FITC-Hy) and FITC-Rhodostomin (FITC-Rn) was measured in both resting and ADP-activated platelets of diluted human platelet-rich plasma. The binding fluorescence of FITC-disintegrins was abolished by EDTA and 7E3, a monoclonal antibody against GPIIb-IIIa. ADP markedly increased the fluorescence intensity of FITC-Tg and FITC-Hy bound on platelets especially when lower doses of these probes were used, whereas it had little effect on that of FITC-Rn. Therefore, FITC-Tg and FITC-Hy can be used for the detection of the activated platelets as noted by a higher ratio of fluorescence intensity (approx. 2-4) between ADP-activated and resting platelets as compared with that (approx. 1-1.3) in the case of FITC-Rn as the probe. The platelets from three patients with Glanzmann's thrombasthenia were probed with FITC-disintegrins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7740464

Liu, C Z; Wang, Y W; Shen, M C; Huang, T F

1994-12-01

263

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

NASA Astrophysics Data System (ADS)

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

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

2013-08-01

264

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

SciTech Connect

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

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

2010-07-02

265

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

E-print Network

1 Silver Staining of 2D Electrophoresis Gels Cécile Lelong, Mireille Chevallet, Sylvie Luche Cedex 9, France 1. Introduction Silver staining of polyacrylamide gels was introduced in 1979 by Switzer staining with Coomassie Blue. However, the first silver staining protocols were not trouble-free. High

Paris-Sud XI, Université de

266

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

Microsoft Academic Search

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

JAMES H. MORRISSEY

1981-01-01

267

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

PubMed Central

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

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

1997-01-01

268

Development of Cell Staining Technique for X-Ray Microscopy  

NASA Astrophysics Data System (ADS)

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.; Liang, K. S.; Yin, G. C.; Chen, F. R.; Je, J. H.; Margaritondo, G.; Hwu, Y.

2007-01-01

269

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

270

Multicolored Stain-free Histopathology with Coherent Raman Imaging  

PubMed Central

Conventional histopathology with hematoxylin & eosin (H&E) has been the gold standard for histopathological diagnosis of a wide range of diseases. However, it is not performed in vivo and requires thin tissue sections obtained after tissue biopsy, which carries risk, particularly in the central nervous system. Here we describe the development of an alternative, multicolored way to visualize tissue in real time through the use of coherent Raman imaging (CRI), without the use of dyes. CRI relies on intrinsic chemical contrast based on vibrational properties of molecules and intrinsic optical sectioning by nonlinear excitation. We demonstrate that multi-color images originating from CH2 and CH3 vibrations of lipids and protein, as well as two-photon absorption of hemoglobin, can be obtained with subcellular resolution from fresh tissue. These stain-free histopathological images show resolutions similar to those obtained by conventional techniques, but do not require tissue fixation, sectioning or staining of the tissue analyzed. PMID:22906986

Freudiger, Christian W.; Pfannl, Rolf; Orringer, Daniel A.; Saar, Brian G.; Ji, Minbiao; Zeng, Qing; Ottoboni, Linda; Ying, Wei; Waeber, Christian; Sims, John. R.; De Jager, Philip L.; Sagher, Oren; Philbert, Martin A.; Xu, Xiaoyin; Kesari, Santosh; Xie, X. Sunney; Young, Geoffrey S.

2013-01-01

271

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

272

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

273

Selection of ovine oocytes by brilliant cresyl blue staining.  

PubMed

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

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

2012-01-01

274

New Giemsa method for the differential staining of sister chromatids  

Microsoft Academic Search

IF human lymphocytes1 or Chinese hamster2 cells are treated with the base analogue 5-bromodeoxyuridine (BrdU) in the latter part of the S period, Giemsa stained chromosomes exhibit a pattern of condensed and extended segments along their length. This phenomenon has been attributed to a delay in the spiralisation pattern of the late replicating regions along the chromosomes. Other experiments3 with

Paul Perry; SHELDON WOLFF

1974-01-01

275

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

276

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

277

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

278

Development of new staining technology "eastern blotting" using monoclonal antibody.  

PubMed

Ginsenosides contained in Panax species were separated by silica gel TLC blotted to a polyvinylidene difluoride (PVDF) membrane which was dipped in a sodium periodide (NaIO(4)) solution and reacted with protein, preparing a ginsenoside-protein conjugate for binding a ginsenoside on a PVDF membrane. The blotted spots were stained by anti-ginsenoside-Rb1 monoclonal antibody (MAb) and anti-ginsenoside-Rg1MAb, respectively. The newly established immunostaining method, eastern blotting was applied for the determination of ginsenosides possessing protopanaxadiol and/or protopanaxatriol. Double staining of eastern blotting for ginsenosides using anti-ginsenoside-Rb1 MAb and anti-ginsenoside-Rg1 MAb promoted complete identification of ginsenosides in Panax species. This technique has been devised for the chromatographic separation and identification of ginsenosides using polyethersulfone (PES) membrane. It caused an acceptable separation of ginsenoside-Rb1, -Rc and -Rd in various ginseng extracts. Newly developed technique is quite simple and applies for immunoassay system. Ginsenosides separated using a PES membrane were directly treated with a NaIO(4) solution and then reacted with bovine serum albumin (BSA) for making a ginsenoside-protein conjugate. After the blocking, anti-ginsenoside-Rb1 MAb recognized a ginsenoside on a PES membrane and then a sec-ond antibody labeled with enzyme reacted to the first antibody. Finally a substrate was oxidized with the enzyme and de-veloped the staining of ginsenosides. The staining spots of ginsenosides on membrane were quantitatively evaluated by NIH Image indicating at least 62.5 ng of each ginsenoside-Rb1, -Rc and -Rd were detected with clarity. The determination range of three ginsenosides was from 0.125 to 2.0 µg of direct amount on PES membrane. PMID:21143134

Morinaga, Osamu; Shoyama, Yukihiro

2011-03-01

279

Validation of image analysis for enzyme histochemical and immunocytochemical staining.  

PubMed

Immunocytochemical and enzyme histochemical analyses of cells and tissues are used to detect changes in the extent of injury and the expression of various molecules. Image analysis quantitation offers an easier, more efficient technique to evaluate these changes. We studied the application of image analysis for evaluating enzyme histochemistry and immunocytochemistry of cells and tissues as a way to assess stroke. Using brain sections, we compared investigator and computer-generated image analysis of 2,3,5-triphenyltetrazolium chloride stained cerebral infarcts in rats subjected to 2 h middle cerebral artery occlusion and 22 h re-perfusion. Both methods documented the infarct volumes with a comparison of means of less then 5%. This suggests no difference between computer- and hand-calculated values. Computer-generated analysis was easier and faster to use. Using endothelial cell monolayers, immunocytochemical staining of a time course of heat shock protein expression was compared to a grading system using fast red chromagen counterstained with hematoxylin. Results demonstrated greater ease and efficiency with computer-generated image analysis compared to other subjective systems of analysis. Image analysis is more useful for detecting small differences in staining, especially when using 3,3-diaminobenzidine as a chromagen. Investigator bias is also reduced using this system. Our comparisons validate the use of this versatile technology to assess more easily both cell and tissues in stroke research. PMID:12503731

Patel, V M; Heinel, L A; Provencio, J J; Vinall, P E; Kramer, M S; Rosenwasser, R H

2002-07-01

280

Study of stained glass window using PIXE PIGE  

NASA Astrophysics Data System (ADS)

We had the opportunity to study a large panel (100 × 80 cm) containing more than 40 stained glass pieces. Among them several come from restorations having taken place at different periods. The study of this rather complex arrangement has been processed by stages: the elemental composition of 16 zones were determined: several differences were identified and among them the Na/K ratio which allowed to set three groups of glass type; the measurement of the Na concentrations by the two techniques give information in bulk (PIGE) and at the near surface (PIXE); the values defined by the ( CPIGE- CPIXE))/ CPIGE plotted in function of the historical estimation of the age of the stained glass pieces (original and restored) indicate a real correlation between the two variables; the red-colored pieces were specially investigated in order to determine which coloration technique was employed (bulk coloration, superficial staining, multilayered flashing, etc.); the corrosion was investigated by scanning two different worsened zones with a 0.5 mm diameter beam spot. This study shows the possibilities of the PIGE-PIXE association, but also points out some weaknesses, which have to be solved by other techniques; unfortunately, in that case, the non-destructive aspect could be lost.

Weber, G.; Vanden Bemden, Y.; Pirotte, M.; Gilbert, B.

2005-10-01

281

Electron microscopy of negatively stained and unstained fibrinogen.  

PubMed

Electron microscope images of negatively stained fibrinogen are predominantly asymmetric rods 450 A in length and about 60 A in width. The molecules appear to have considerable flexibility, and mass distribution along the major axis is not uniquely distinguished despite apparent beading in some particles. Scanning transmission electron microscopy of unstained fibrinogen again demonstrates that a majority of molecules are rodlike. The results differ from those obtained by negative staining in that a substantial fraction of images are trinodular with striking resemblance to those obtained by C. E. Hall and H. S. Slayter [J. Biophys. Biochem. Cytol. (1959) 5, 11--16] using the mica replica technique. The above results were obtained on glow-discharged carbon substrate films by a simple low-concentration, long-attachment-time modification of standard deposition methods that is diffusion controlled and depends on concentration and time but is independent of pH, buffer, and other staining conditions. Evidence is presented that standard attachment procedures result in artifactual images. Any models of fibrinogen in solution consequently must encompass properties that permit its visualization as an asymmetetric rod by electron microscopy as first suggested by Hall and Slayter 20 years ago. PMID:6158041

Estis, L F; Haschemeyer, R H

1980-06-01

282

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

PubMed

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

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

2009-05-01

283

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

NASA Astrophysics Data System (ADS)

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

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

2005-02-01

284

Machine vision system for automated detection of stained pistachio nuts  

NASA Astrophysics Data System (ADS)

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

Pearson, Tom C.

1995-01-01

285

Stains and smears: resident guide to bedside diagnostic testing.  

PubMed

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

Bronfenbrener, Roman

2014-12-01

286

Early localized morphea mimicking an acquired port-wine stain.  

PubMed

Port-wine stains (PWS) and morphea are distinct conditions that are easily recognized and diagnosed in pediatric patients. Rarely, early localized morphea may mimic an acquired PWS. We present 4 such cases, in two of which the initial clinical impression of acquired PWS was thought to be confirmed by histopathology. A diagnosis of morphea was made approximately 6 months to 3 years after the onset of the acquired PWS. Clinicians should be aware that an apparent acquired PWS may be an early manifestation of localized morphea and continue to monitor these lesions. PMID:20850196

Nijhawan, Rajiv I; Bard, Susan; Blyumin, Marianna; Smidt, Aimee C; Chamlin, Sarah L; Connelly, Elizabeth A

2011-04-01

287

Development of an N-hydroxysuccinimidyl fluorescein-O-acetate-labeled probe for competitive capillary electrophoretic immunoassay of bovine serum albumin.  

PubMed

A highly fluorescent reagent, N-hydroxysuccinimidyl fluorescein-O-acetate (SIFA) has been labeled on bovine serum albumin (BSA) to construct a new immunofluorescent probe, SIFA-BSA, for the competitive immunoassay of BSA with capillary electrophoresis. Labeling conditions of SIFA with BSA such as the reaction molar ratio and reaction time, as well as separation conditions for free and antibody-bound SIFA-BSA including the pH and concentration of buffer were systematically investigated. Under the optimized conditions, SIFA-labeled BSA and free BSA competitively reacted with a limited amount of anti-BSA antibody. Free and antibody-bound SIFA-BSA could be well separated within 5min using 100mM boric acid buffer (pH 9.3) for background electrolyte, 28kV for the separation voltage, and 25 degrees C for the column temperature. With the proposed method, a much lower detection limit (S/N=3) of 0.1microg/mL was obtained compared with the competitive capillary electrophoresis immunoassay of BSA with fluorescein isothiocyanate (FITC) labeled BSA probe, in which the detection limit (S/N=3) is 0.0031mg/mL. PMID:20441977

Deng, Ying-Hua; Zhang, Zi-Xing; Zhang, Hua-Shan; Wang, Hong

2010-06-15

288

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

PubMed

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

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

2014-10-01

289

Investigation of influence of different values of pH on mechanisms of binding of human serum albumin with markers of fluorescein family  

NASA Astrophysics Data System (ADS)

Objective and background dataThis work is dedicated to investigation of influence of different values of pH on mechanisms of binding of human serum albumin (HSA) with markers of fluorescent family - eosin, erythrosin and fluorescein. For this purpose were detected changes in markers fluorescence, in markers molecular association, in the effective constants of binding of markers to HSA and also in changes in related chemical bonds in HSA-marker association. Such analysis of changes in binding of biomolecules (such as proteins) with different ligands (such as markers) is extremely interesting from the point of view of a biomedicine and pharmaceuticals, so from the point of view of bionanotechnology: for example, at creation of new drugs. MethodsThe investigations of steady-state fluorescence, polarized fluorescence, molecular association of markers of fluorescein family in HSA solutions are presented in this work, also the analysis of changes in related chemical bonds in HSA-marker association by Raman spectroscopy is done, and also the effective constants of binding of markers to HSA are calculated. Results and conclusionAll investigations show the leading role of chemical and electrostatic interactions between markers and HSA. The received data allow one to get information about mechanisms of interaction of markers to HSA, that can be useful at research of structure and properties of binding Centers (drug-binding Centers) of transport blood protein-HSA, what is of great importance in medical investigations of binding of drugs to HSA.

Vlasova, Irina M.; Saletsky, Alexander M.

2009-11-01

290

Detection of embolization of vessels by a double staining technique.  

PubMed

Air seeding and water vapor embolization of vessels were induced in branches of Acer rubrum by freezing-thawing experiments. A technique has been developed for detecting embolized vessels at the microscopical level. In this technique, one dye is used to label functioning vessels prior to freezing while the second dye is offered after a freezing-thawing cycle at vacuum pressure or at atmospheric pressure in order to reveal water vapor embolization or recovering at 1 bar. Double staining is achieved by differential staining of vessel walls and of their particular contact pits facing paratracheal contact cells. The water potential of the leaves and the rate of uptake of the dye solutions was recorded during the experiments using a Scholander pressure bomb and a potetometer, respectively. While almost 100% of the vessels proved to be functioning in the outer 5 to 7 annual rings, one freezing-thawing cycle inhibited water conduction in up to 82 to 98% of the vessels provided vacuum (0.13 bar) was applied. Rapid recovering however is observed, if the second dye is introduced at atmospheric pressure. PMID:23195303

Sauter, J J

1984-01-01

291

Analysis of surface stains on modern gold coins  

NASA Astrophysics Data System (ADS)

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

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

2013-07-01

292

[Sensitivity of various study technics in blood stains].  

PubMed

Quadratic pieces of fleece measuring 16 mm2 were soaked with 10 different blood-samples in the dilution steps of 1:1, 1:10, 1:100, 1:1000, respectively, and were tested in blood group typing and identification tests of forensic serology. The above spezified dilutions correspond with 5 microliters, 0.5 microliter, 0.05 microliter and 0.005 microliter of blood, respectively. The detection limit of the microspectrometric test for blood was the dilution 1:10, of the porphyrine test a dilution above 1:100, whereas the preliminary test for blood (peroxidase) succeeded always up to a dilution of 1:1000 and the species determination by the radial immunodiffusion test in agar gels succeeded in most cases op to a dilution of 1:1000. The detection limit of the anti-human globulin inhibition test was between the dilution steps 1:10 and 1:100 when non-titrated and undiluted anti-human globulin serum was used. Gc- and ABO-grouping were possible up to a dilution of 1:100 and were thus the most sensitive grouping systems. Phenotyping of the enzyme-systems and the Gm/Km-system usually required stains with considerably higher blood concentrations i.e. stains of undiluted blood. PMID:2390000

Förster, R; Bruder, W

1990-01-01

293

In vivo corneal and conjunctival epithelial nuclear stain.  

PubMed

Epithelium plays a very important structural and functional role in the cornea and conjunctiva. Evaluation of the epithelium is the first step in diagnosing many pathologic states. We have developed a new technique for the in vivo staining of nuclei of corneal and conjunctival epithelium in rabbits and guinea pigs. Several drops of 0.01%, 0.1%, 0.25%, 0.5%, or 1.0% toluidine blue or 1.0% methylene blue were applied to the conjunctival sac to stain epithelial cells. The cells picked up the vital dye within 5 minutes and could be photographed at 30X with the Keeler-Konan wide-field specular microscope. Cells and nuclei were clearly observable. Photographs could be further enlarged to enhance details. Wash out time was rapid and no toxic effects were observed. This technique adds a new dimension to the study of epithelium in normal and pathologic states in experimental animals. This technique may also be applicable to human eyes for discerning such diseases as carcinoma, herpes simplex, or superior limbic keratoconjunctivitis. PMID:2419030

Wander, A H; Neumeister, R D; Tschismadia, I; Choromokos, E A; Masukawa, T

294

A Comparison of Heat versus Methanol Fixation for Gram Staining Bacteria  

ERIC Educational Resources Information Center

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

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

2009-01-01

295

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

Microsoft Academic Search

Previous bacterial transport studies have utilized fluorophores which have been shown to adversely affect the physiology of stained cells. This research was undertaken to identify alternative fluorescent stains that do not adversely affect the transport or viability of bacteria. Initial work was performed with a groundwater isolate, Comamonas sp. strain DA001. Potential compounds were first screened to determine staining efficiencies

MARK E. FULLER; SHERYL H. STREGER; RANDI K. ROTHMEL; BRIAN J. MAILLOUX; JAMES A. HALL; TULLIS C. ONSTOTT; JAMES K. FREDRICKSON; DAVID L. BALKWILL; MARY F. DEFLAUN

2000-01-01

296

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

E-print Network

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

Hubel, David

297

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

Microsoft Academic Search

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

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

1996-01-01

298

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

Microsoft Academic Search

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

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

2000-01-01

299

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

Microsoft Academic Search

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

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

1997-01-01

300

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

Technology Transfer Automated Retrieval System (TEKTRAN)

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

301

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

E-print Network

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

Lebendiker, Mario

302

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

E-print Network

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

Lebendiker, Mario

303

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

PubMed Central

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

Ankle, Madhuri R; Joshi, Priya S

2011-01-01

304

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

E-print Network

of buffelgrass (Cenchrus c///ar/s L. cv. 'Common' ), 'T-704' buffelgrass (C. c///ar/s Pl 409704), birdwoodgrass (C. ser/gervs Vahl. Pl 193444), pearl millet [Penn/se/um g/aucvm (L. ) R. Br. cv. 'Tift 23DB'], and P. or/en/a/e Rich. cv. 'Cowboy' was used... mM Ca, 2 mM B, and 10 g L' gum agar appeared acceptable for estimating pollen viability for all taxa. In vitro germination, fluorescein diacetate (FDA), and iodine staining were then evaluated on pollen of birdwoodgrass and pearl millet. In vitro...

Hill, Jerry Leon

1991-01-01

305

Pulsed photothermal radiometry of port-wine-stain lesions.  

PubMed

Pulsed photothermal radiometry is used to map the heat deposition in human skin after a short laser pulse. It uses an IR (HgCdTe) detector for a rapid noncontact measurement of the skin surface temperature based on the blackbody emission in the 8-12-microm spectrum. The heat deposited by the laser pulse in the superficial epidermis causes an immediate temperature jump, and the heat deposited in basal epidermal melanin and deep port wine stains diffuses to the surface before detection. The time course of the surface temperature T(z = 0, t), indicates the initial spatial distribution of heat, T(z, t = 0), deposited by the laser. PMID:20820403

Jacques, S L; Nelson, J S; Wright, W H; Milner, T E

1993-05-01

306

Comparison of Cleaning Methods for Stained Glass Windows  

NASA Astrophysics Data System (ADS)

Any cleaning process for stained glass windows has to consider the effectiveness of the treatment but also the potential damage for the art object. A variety of mechanical and chemical methods is currently used in restoration practice. The most effective ones are criticized because of their long-term risks. Therefore, an interdisciplinary research project, carried out in Germany and funded by the "Deutsche Bundesstiftung Umwelt (DBU)" had explored the possibilities and limits of lasers for cleaning glass windows. At previous LACONA conferences the Excimer Laser equipment and results from the research project have been presented. This contribution puts the cleaning experiments in a broader context, by comparing lasers with conventional techniques. Scientific and practical aspects will be discussed, focussing on the removal of crust and aged polymers.

Römich, H.; Mottner, P.; Hildenhagen, J.; Dickmann, K.; Hettinger, G.; Bornschein, F.

307

Exposure to lead in stained glass work. An environmental evaluation.  

PubMed

An environmental evaluation was conducted to determine lead exposure in a group of crafts people who produce stained glass and Tiffany glass. The environmental evaluation consisted of air sampling for potential lead emissions from solder and of work area dusts. In addition, the completion of a questionnaire, observation of work practices and noting of other details relevant to hazardous exposures were carried out. Lead concentrations in air were found to be well below the ACGIH TLV-TWA of 150 micrograms/m3. High lead concentrations were found in the work area dust samples. Exposure to high concentrations of lead could occur by ingestion as a result of neglect of basic hygiene precautions. PMID:8178119

Pant, B C; Harrison, J R; Long, G W; Gupta, S

1994-01-25

308

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

309

Conjugates of a Photoactivated Rhodamine with Biopolymers for Cell Staining  

PubMed Central

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

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

2014-01-01

310

Classification and naming of dyes, stains and fluorochromes.  

PubMed

A classification of dyes and other colorants is proposed, based on the chemical features responsible for their visibility and generally consonant with the writings of modern color chemists. The scheme differs in several respects from that of the Colour Index (CI), but it retains some traditional small groups of dyes that include biological stains. Natural dyes, recognized as a group in the CI, are placed with or near synthetic dyes with identical or similar chromophores. The new scheme also provides categories for dyes and fluorochromes that do not have places in the CI classification. Some CI categories, including lactones, aminoketones and hydroxyketones, are not recognized in this new scheme, which is adopted in the forthcoming 10th edition of Conn's Biological Stains: a Handbook of Dyes and Fluorochromes for Use in Biology and Medicine. Some rules are also set out for the spelling of trivial names, which has long been inconsistent in scientific literature. The ending '-ine' is used for compounds derived from organic bases (e.g., fuchsine and thionine, not fuchsin or thionin), and names ending in '-in' are for compounds that are not bases or their derivatives (e.g., eosin and phloxin, not eosine or phloxine). Initial capital letters are used only for words that are names of people or places (e.g., Nile blue or Congo red) and for the 'generic' components of CI application names (as in Acid yellow 36). Other words, including trade names that have fallen into common usage are not capitalized (e.g., alcian blue, biebrich scarlet, coomassie blue). The recommended spellings of some dyes differ from those commonly seen in vendors' catalogs and in biological publications, but they are generally consistent with English and American dictionaries, with recent writings in English by color chemists, and with the trivial names of other organic compounds. PMID:11871748

Kiernan, J A

2001-01-01

311

PREFERENTIAL STAINING OF NUCLEIC ACID-CONTAINING STRUCTURES FOR ELECTRON MICROSCOPY  

PubMed Central

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

Huxley, H. E.; Zubay, G.

1961-01-01

312

Argyrophilic nucleolar organiser region (AgNOR) staining in normal bone marrow cells.  

PubMed Central

Fifteen normal bone marrow aspirates were stained with the agyrophilic nucleolar organiser region (AgNOR) method. The results of the specific staining AgNORs as well as nuclear and cytoplasmic staining were analysed. A system was devised to characterise precisely the AgNORs present in the nuclei of bone marrow cells. Particular types of bone marrow cells had a characteristic AgNOR and non-AgNOR staining pattern. The bone marrow cells were identified easily and reliably with AgNOR staining and the method was especially useful for lymphocytes, plasma cells, erythroid cells, basophils/mast cells, monocytes and cells containing haemosiderin. The immature haemopoietic cells exhibited more and larger AgNORs than the more mature cells. It is concluded that AgNOR staining can be used to study bone marrow cells by providing additional information when used in conjunction with conventional stains. Images PMID:1698824

Nikicicz, E P; Norback, D H

1990-01-01

313

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. PMID:24825573

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

2014-08-01

314

Fluorescent stain method for the simultaneous determination of mitochondrial potential and integrity of plasma and acrosomal membranes in boar sperm.  

PubMed

The purpose of this study was to validate a technique for simultaneous evaluation of the plasma, acrosomal and mitochondrial membranes in boar spermatozoa, using an association of fluorescent probes: Propidium iodide (PI), fluorescein isothiocyanate-conjugated Pisum sativum agglutinin (FITC-PSA) and JC-1. Three ejaculates from each of four different boars, all showing motility >or=80% and abnormal morphology stain technique. To a 150-microl aliquot of diluted semen it was added 3 microl of PI (0.5 mg/ml), 2 microl of JC-1 (153 microm) and 50 microl of FITC-PSA (100 microg/ml). Samples were incubated at 38.5 degrees C for 8 min, in the dark. An 8-microl sample was put on a slide, coverslipped and immediately evaluated by epifluorescent microscopy. The association of fluorescent probes was divided into eight cell classes, according to plasma membrane integrity, intact acrosome and mitochondrial function. For plasma membrane integrity, detected by PI probe, the equation: (p < 0.0001) and R(2) = 0.97 was obtained. The intact acrosome, verified by the FITC-PSA probe, produced the equation: (p < 0.0001) and R(2) = 0.98. The mitochondrial potential, marked by JC-1, was estimated by the equation: (p < 0.001) and R(2) = 0.99. The group of spermatozoa with combined intact plasma membrane, intact acrosome and high mitochondrial potential (IPIAH), was estimated by the equation: (p < 0.0001) and R(2) = 0.97. The resulting linear equations demonstrate that this technique is efficient and practical for the simultaneous evaluations of the plasma, acrosomal and mitochondrial membranes in boar spermatozoa. PMID:17348977

de Andrade, A F C; de Arruda, R P; Celeghini, E C C; Nascimento, J; Martins, S M M K; Raphael, C F; Moretti, A S

2007-04-01

315

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

PubMed

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

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

2015-01-01

316

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. PMID:8809467

Woods, G L; Walker, D H

1996-01-01

317

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

PubMed Central

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

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

2015-01-01

318

Is there an optimal laser treatment for port wine stains?  

PubMed

Optimization of laser treatment of port wine stains (PWS) is discussed from the standpoint of heat production. Laser wavelength, irradiation time, heat conduction, and external epidermal cooling are the variables considered in conjunction with absorbing and scattering behavior of a PWS-model consisting of epidermis, dermis, and ectatic blood vessel. Ideal treatment is defined as minimal heating of the epidermis and upper dermis, but with irreversible damage to the capillary wall. The analysis shows that irradiation times of 1-10 ms in conjunction with external epidermal cooling may give optimal results. The wavelength of choice is 577 nm, followed by 540, 415, 560, and 500 nm (argon laser). The ruby and Nd-YAG lasers are predicted to damage the epidermis and dermis at all times when the capillary is coagulated. Concurrent cooling to prevent epidermal-dermal damage is also recommended here. The CO2 laser is predicted to be the worst laser and, according to our analysis, should not be used to treat PWS. Both upper dermal and capillary destruction can only result from heat conduction from the damaged epidermis and external cooling cannot be applied here. PMID:3959719

van Gemert, M J; Welch, A J; Amin, A P

1986-01-01

319

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

320

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

321

The challenges of analysing blood stains with hyperspectral imaging  

NASA Astrophysics Data System (ADS)

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

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

2014-06-01

322

p63 immunohistochemical staining is limited in soft tissue tumors.  

PubMed

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

Jo, Vickie Y; Fletcher, Christopher D M

2011-11-01

323

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

Microsoft Academic Search

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

Masahiro Sakagami; Wataru Kinoshita; Kiyoyuki Sakon; Yuji Makino

2003-01-01

324

Fluorescein angiographic evaluation of the effect of latanoprost treatment on blood-retinal barrier integrity: A review of studies conducted on pseudophakic glaucoma patients and on phakic and aphakic monkeys  

Microsoft Academic Search

Endogenous prostaglandins (PGs) have been claimed to play a role in the development of cystoid macular edema (CME). Two fluorescein angiographic studies evaluating the effect of latanoprost, a new ocular hypotensive PG analogue, on blood-retinal barrier integrity are, therefore, reviewed here. In the first study, six of eight unilaterally aphakic cynomolgus monkeys were treated bilaterally once daily for six months

Philip F. J. Hoyng; Alexander H. Rulo; Erik L. Greve; Maria Astin; M. Gjötterberg

1997-01-01

325

Hematoxylin Staining Reveals a Decrease in Nucleolar Diameter of Pig Oocytes Before Germinal Vesicle Breakdown  

PubMed Central

During oocyte growth, the morphology of the nucleolus changes into a compact and homogenous structure. The compact nucleoli in full-grown oocytes are not stained by aceto-orcein staining or immunofluorescence staining. In this study, we developed a hematoxylin staining method for pig oocytes in whole-mount preparations to visualize the nucleoli. Nucleoli of growing and full-grown oocytes were stained blue with hematoxylin. Using this staining method, the changes in the oocyte nucleolus during maturation were examined. The nucleolar diameter gradually decreased in maturing oocytes (10.7 ± 0.1 ?m to 9.0 ± 0.7 ?m, P<0.05) before germinal vesicle breakdown (GVBD). The results suggest that the nucleolar volume of oocytes decreases before GVBD. PMID:23856597

Nobata, Tadatoshi; Kyougoku, Hirohisa; Miyano, Takashi

2013-01-01

326

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

NASA Technical Reports Server (NTRS)

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

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

2002-01-01

327

Identification of mast cells in bronchoalveolar lavage fluid. Comparison between different fixation and staining methods.  

PubMed

The present study was carried out to compare the effectiveness of different fixation and staining methods in the identification of mast cells obtained by bronchoalveolar lavage from patients with interstitial lung diseases. Cell preparations were fixed with formaldehyde or methanol. Mast cells were identified by metachromatic staining with May Grünwald Giemsa, Toluidine blue or Gallamine blue Giemsa. After formaldehyde fixation only a few mast cells were identified, regardless of the stain method used. In contrast, a significantly higher number of mast cells were observed after methanol fixation. Using this fixative, Toluidine blue stain showed a higher number of mast cells than May Grünwald Giemsa. However, there was no difference between Toluidine and Gallamine blue Giemsa in the number of cells observed. The easy identification of mast cells after staining with toluidine, combined with its easy application, suggest that Toluidine blue stain after methanol fixation is the most useful method for determining the presence of mast cells in lavage fluid. PMID:1711798

Xaubet, A; Moisés, J A; Agustí, C; Martos, J A; Picado, C

1991-04-01

328

Double-Staining Method for Differentiation of Morphological Changes and Membrane Integrity of Campylobacter coli Cells  

PubMed Central

We developed a double-staining procedure involving NanoOrange dye (Molecular Probes, Eugene, Oreg.) and membrane integrity stains (LIVE/DEAD BacLight kit; Molecular Probes) to show the morphological and membrane integrity changes of Campylobacter coli cells during growth. The conversion from a spiral to a coccoid morphology via intermediary forms and the membrane integrity changes of the C. coli cells can be detected with the double-staining procedure. Our data indicate that young or actively growing cells are mainly spiral shaped (green-stained cells), but older cells undergo a degenerative change to coccoid forms (red-stained cells). Club-shaped transition cell forms were observed with NanoOrange stain. Chlorinated drinking water affected the viability but not the morphology of C. coli cells. PMID:12324366

Alonso, Jose L.; Mascellaro, Salvatore; Moreno, Yolanda; Ferrús, María A.; Hernández, Javier

2002-01-01

329

Double-staining method for differentiation of morphological changes and membrane integrity of Campylobacter coli cells.  

PubMed

We developed a double-staining procedure involving NanoOrange dye (Molecular Probes, Eugene, Oreg.) and membrane integrity stains (LIVE/DEAD BacLight kit; Molecular Probes) to show the morphological and membrane integrity changes of Campylobacter coli cells during growth. The conversion from a spiral to a coccoid morphology via intermediary forms and the membrane integrity changes of the C. coli cells can be detected with the double-staining procedure. Our data indicate that young or actively growing cells are mainly spiral shaped (green-stained cells), but older cells undergo a degenerative change to coccoid forms (red-stained cells). Club-shaped transition cell forms were observed with NanoOrange stain. Chlorinated drinking water affected the viability but not the morphology of C. coli cells. PMID:12324366

Alonso, Jose L; Mascellaro, Salvatore; Moreno, Yolanda; Ferrús, María A; Hernández, Javier

2002-10-01

330

Staining plant cells with silver. I. The salt-nylon technique.  

PubMed

A technique is described for selectively silver staining nucleoli, active nucleolus organizers, nucleolar material attached to chromosomes, kinetochores, synaptonemal complexes, and chromosome cores in plant cells. The technique, called salt-nylon silver staining, involves spreading cells on glass slides, treating the cells with a solution of saline sodium citrate, and incubating the cells in a silver nitrate solution covered with nylon screen. Selected variables important for achieving reliable silver staining are considered. PMID:1714766

Stack, S; Herickhoff, L; Sherman, J; Anderson, L

1991-01-01

331

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

Microsoft Academic Search

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

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

1992-01-01

332

Staining paraffin embedded sections of scald of barley before paraffin removal.  

PubMed

Staining of paraffin embedded sections with periodic acid-Schiff reagent and fast green before paraffin removal resulted in differentiation of barley seed and leaf tissue from fungal structures of Rhynchosporium secalis. Crystal violet, toluidine blue O and antiline blue also successfully stained fungal structures of R. secalis in barley leaf tissues. Staining of embedded sections before paraffin removal allows simple processing of a series of sections, saves time and reduces solvent consumption. PMID:9290905

Xi, K; Burnett, P A

1997-07-01

333

Multispectral image enhancement for H&E stained pathological tissue specimens  

NASA Astrophysics Data System (ADS)

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

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

2008-03-01

334

Banding patterns in rat incisor enamel stained by histochemical complexing methods for calcium.  

PubMed

A characteristic banding pattern can be visualized at the surface of the rat incisor in the maturation zone of amelogenesis by staining with glyoxal bis(2-hydroxyanil) (GBHA). Other banding patterns can be obtained with certain histological and fluorochrome stains and by radioautography following 45Ca injection. In this study, several histochemical reagents known to complex with different states of calcium were used to stain the surface of enamel. Rat incisors were quickly dissected and immediately immersed in solutions containing the following calcium-binding reagents: arsenazo III, calmagite, murexide, N,N-naphthaloylhydroxylamine, and calcein. Routinely, one contralateral lower incisor from each pair was counterstained with GBHA in order to relate each of the staining patterns to the banded distribution of maturation ameloblasts that is reflected by the characteristic GBHA staining pattern in the enamel. Each of the reagents used in this study demonstrated a staining pattern consisting of a series of broad bands running transversely and obliquely across the enamel. In all cases, the dyes stained predominantly that enamel associated with ruffle-ended ameloblasts, i.e. enamel left unstained by GBHA. Some of the reagents also stained enamel in the secretion zone. The appearance and distribution of the staining patterns reflect the banded distribution of maturation ameloblasts and appear to be controlled on a time scale related to the rapid modulation of these cells. PMID:2471424

McKee, M D; Warshawsky, H

1989-05-01

335

Silver staining of proteins in polyacrylamide gels: a general overview Thierry Rabilloud*, Laurent Vuillard+ , Claudine Gilly and Jean Jacques Lawrence  

E-print Network

1 Silver staining of proteins in polyacrylamide gels: a general overview Thierry Rabilloud (Running title): overview of silver staining methods for proteins +: Institut Laue-Langevin , BP156, 38042 silver staining of proteins, which are recalled in this paper, several methods of silver staining

Boyer, Edmond

336

Effects of N-methyl pyrrolidone on the uptake of hypericin in human bladder carcinoma and co-staining with DAPI investigated by confocal microscopy.  

PubMed

Photodynamic diagnosis (PDD) using hypericin (HY), a natural photosensitizer, detects bladder cancer significantly better than white light endoscopy. However, the lipophilicity of HY complicates its administration for clinical applications. Currently, pharmaceutical preparations for HY without plasma protein are being developed. Formulations containing a biocompatible solvent, N-methyl pyrrolidone (NMP) have been shown to enhance the photodynamic therapeutic effects of HY. It was recently reported that, NMP formulations of HY were able to produce significantly higher contrast for fluorescence detection of tumors than albumin-containing HY formulations. This present work hypothesizes that NMP acts both as a solvent and penetration enhancer to improve the delivery of HY into cells by increasing the permeability of cell membranes. This paper reports the use of 3-D confocal microscopy to monitor real-time uptake of HY in human carcinoma. 3-D confocal microscopy was used to investigate the possibility of nuclear localization of HY in MGH cells. The fluorescence of HY was confirmed to be emitted from HY containing cells using spectrometry. The localization of a DNA fluorescent probe 4', 6-diamidino-2-phenylindole dihydrochloride (DAPI) was used to confirm the possibility of colocalization of DAPI and HY. The colocalization analysis in the present study suggests that it was very unlikely that HY colocalized in the nucleus that was stained by DAPI. Fluorescein leakage tests showed that 1% NMP changes the permeability of cell membranes, and enhanced the delivery of HY into cells resulting in lower cell survival ratios. Thus, NMP was able to enhance the photodynamic therapeutic effects of HY on cancer cells. PMID:17877426

Saw, Constance Lay Lay; Olivo, Malini; Wohland, Thorsten; Fu, Chit Yaw; Kho, Kiang Wei; Soo, Khee Chee; Sia Heng, Paul Wan

2007-10-01

337

The effectiveness of two different battery-powered toothbrushes on whitening through removal of stain.  

PubMed

Whitening dentifrices have recently become popular, achieving a whitening benefit by surface chemical action and/or abrasion that serve to remove extrinsic stains and/or prevent extrinsic stain buildup. Some powered toothbrushes have also been demonstrated to have whitening/stain-removal efficacy when used with standard dentifrice products. The objective of this study was to evaluate the effectiveness of an experimental prototype powered toothbrush (Crest SpinBrush Pro) and a commercially available powered toothbrush (Crest SpinBrush) on dental whitening through the removal of extrinsic stain. This study was a randomized, controlled, examiner-blind, parallel-group design, which examined extrinsic stain removal over four weeks of brushing by 70 subjects. Following a three-week period of stain induction using rinses of chlorhexidine and tea, subjects were randomized to use one of the two toothbrushes. Tooth stain was scored using the Lobene stain index at baseline, and after two and four weeks of toothbrush use. Prior to statistical analysis, the stain scores were averaged on a per-subject basis. The stain reductions (baseline minus post-treatment) in average scores were calculated and analyzed using an analysis of covariance, with baseline average score as the covariate. The experimental prototype powered toothbrush (Crest SpinBrush Pro) delivered statistically significant (p < 0.001) adjusted (via analysis of covariance) mean reductions from baseline in Lobene composite stain scores of 1.90 and 2.11 after 2 weeks and 4 weeks of use, respectively. These results represent 56-62% reductions in extrinsic stain compared to baseline. The commercially available powered toothbrush (Crest SpinBrush) also delivered statistically significant (p < 0.001) adjusted (via analysis of covariance) mean reductions from baseline in Lobene composite stain scores, with magnitudes of 1.95 and 2.22 after two and four weeks of use, respectively. These results represent 60-68% reductions in extrinsic stain compared to baseline. In conclusion, both the experimental prototype powered toothbrush and the commercially available powered toothbrush were effective in removing extrinsic tooth stain. PMID:12518493

Karpinia, Katherine; Magnusson, Ingvar; Biesbrock, Aaron R; Walters, Patricia A; Bartizek, Robert D

2002-01-01

338

Evaporation stains: suppressing the coffee-ring effect by contact angle hysteresis.  

PubMed

A ring-shaped stain is frequently left on a substrate by a drying drop containing colloids as a result of contact line pinning and outward flow. In this work, however, different patterns are observed for drying drops containing small solutes or polymers on various hydrophilic substrates. Depending on the surface activity of solutes and the contact angle hysteresis (CAH) of substrates, the pattern of the evaporation stain varies, including a concentrated stain, a ringlike deposit, and a combined structure. For small surface-inactive solutes, the concentrated stain is formed on substrates with weak CAH, for example, copper sulfate solution on silica glass. On the contrary, a ringlike deposit is developed on substrates with strong CAH, for example, a copper sulfate solution on graphite. For surface-active solutes, however, the wetting property can be significantly altered and the ringlike stain is always visible, for example, Brij-35 solution on polycarbonate. For a mixture of surface-active and surface-inactive solutes, a combined pattern of a ringlike and concentrated stain can appear. For various polymer solutions on polycarbonate, similar results are observed. Concentrated stains are formed for weak CAH such as sodium polysulfonate, and ring-shaped patterns are developed for strong CAH such as poly(vinyl pyrrolidone). The stain pattern is actually determined by the competition between the time scales associated with contact line retreat and solute precipitation. The suppression of the coffee-ring effect can thus be acquired by the control of CAH. PMID:23721254

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

2013-06-25

339

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

Code of Federal Regulations, 2012 CFR

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

2012-01-01

340

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

Code of Federal Regulations, 2014 CFR

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

2014-01-01

341

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

Code of Federal Regulations, 2013 CFR

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

2013-01-01

342

Nuclear staining and culture requirements for in vitro maturation of domestic bitch oocytes  

Microsoft Academic Search

Oocyte nuclear staining and culture requirements for in vitro maturation (IVM) in the bitch have yet to be fully investigated. In the first part of this study we investigated 7 methods for labeling nuclear material (573 oocytes). The most favorable method involved fixation plus aceto-orcein staining and light microscopy. The influence of serum supplementation of the culture medium for IVM

D. A Hewitt; P. F Watson; G. C. W England

1998-01-01

343

A simplified method for differential staining of aborted and non-aborted pollen grains  

Technology Transfer Automated Retrieval System (TEKTRAN)

The ability to use chemical staining to discriminate aborted from non-aborted pollen grains has well-known practical applications in agriculture. A commonly used technique for assessing pollen vitality, Alexander’s stain, uses chloral hydrate, phenol and mercuric chloride, all of which are highly to...

344

Combined histochemical staining, RNA amplification, regional, and single cell cDNA analysis within the hippocampus  

Microsoft Academic Search

The use of five histochemical stains (cresyl violet, thionin, hematoxylin & eosin, silver stain, and acridine orange) was evaluated in combination with an expression profiling paradigm that included regional and single cell analyses within the hippocampus of post-mortem human brains and adult mice. Adjacent serial sections of human and mouse hippocampus were labeled by histochemistry or neurofilament immunocytochemistry. These tissue

Stephen D Ginsberg; Shaoli Che

2004-01-01

345

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

E-print Network

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

Paris-Sud XI, Université de

346

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

PubMed

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

Dorri, Yaser; Kurien, Biji T

2010-09-15

347

Fading of auramine-stained mycobacterial smears and implications for external quality assurance.  

PubMed

Light-emitting diode fluorescence microscopy is being scaled up for tuberculosis control, but fading of auramine-stained slides could compromise external quality assurance. We stored auramine-stained slides and reexamined them over time. Slides stored in all environments faded quickly, with significant changes in the proportion of positive slides in as little as 1 week. PMID:21430105

Minion, Jessica; Shenai, Shubhada; Vadwai, Viral; Tipnis, Tejashree; Greenaway, Christina; Menzies, Dick; Ramsay, Andrew; Rodrigues, Camilla; Pai, Madhukar

2011-05-01

348

Analysis of Staining Observed on Structures in the Georgetown, South Carolina Area  

SciTech Connect

Beginning around 1970, the Georgetown, SC, community complained about black dust and red stains collecting on houses, cars, boats, and other structures. The community, through the South Carolina Department of Health and Environmental Control (SCDHEC), seeks to identify the source or cause of the staining and ways to reduce or eliminate it in the future.

Cramer, Stephen D.; Covino, Bernard S. Jr.; Govier, R. Dale

2002-05-01

349

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

Microsoft Academic Search

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

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

1988-01-01

350

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

PubMed Central

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

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

2014-01-01

351

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

NASA Astrophysics Data System (ADS)

Yellow decoration effects in stained glasses using silver staining were first applied in the beginning of the 14th century. The glass piece being decorated was usually painted on its side intended to be facing the exterior environment, and then fired to temperatures between 500 and 650 °C, resulting in colours ranging from pale lemon to deep orange. Stained glass fragments painted by this process and belonging to the Convento de Cristo, in Tomar, Portugal, were characterised using micro-PIXE, and complemented with other analytical techniques, namely UV-Vis spectroscopy and XRF. Preliminary analysis showed that a mixture of Ag and Cu was used for the production of the yellow staining. In order to understand this staining process and the influence of the firing temperature on the resulting colours, several soda and potash glasses with compositions similar to those of medieval glasses were produced and characterised. The role played by the addition of Cu in the final colours was also investigated.

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

2011-10-01

352

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

PubMed

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

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

2012-11-01

353

A highly sensitive solid substrate room temperature phosphorimetry for carbaryl detection based on its activating effect on NaIO4 oxidizing fluorescein.  

PubMed

Fluorescein (HFin) could emit strong and stable room temperature phosphorescence (RTP) signal on polyamide membrane (PAM) using Pb(2+) as the ion perturber. Carbaryl could activate effect on NaIO4 oxidating HFin, which caused the RTP signal of the system to quench sharply. The phosphorescence intensity (?I p) of activating system higher 3.3 times (119.4/36.0) than that of non-activating system, and is directly proportional to the content of carbaryl. Thus, an activating solid substrate room temperature phosphorimetry (SSRTP) for carbaryl detection has been established. This sensitive (the limit of quantification (LOQ) was 2.0?×?10(-13) g mL(-1)), selective, simple and rapid method has been applied to determine trace carbaryl in water samples with the results consisting with those obtained by fluorimetry, showing its high accuracy. The apparent activation energy (E) and rate constant (k) of this activating reaction were 20.77 kJ mol(-1) and 1.85?×?10(-4) s(-1), respectively. Meanwhile, the mechanism of activating SSRTP for carbaryl detection was also discussed using infrared spectra (IR). PMID:25294182

Liu, Jiaming; Huang, Qitong; Liu, Zhen-bo; Lin, Xiaofeng; Zhang, Li-Hong; Lin, Chang-Qing; Zheng, Zhi-Yong

2014-11-01

354

Reduced Fluorescein Angiography and Fundus Photography Use in the Management of Neovascular Macular Degeneration and Macular Edema During the Past Decade  

PubMed Central

Purpose. We assessed recent trends in the use of diagnostic testing for neovascular age-related macular degeneration (NVAMD) and macular edema (ME). Methods. Claims data from a managed-care network were analyzed on patients with NVAMD (n = 22,954) or ME (n = 31,810) to assess the use of fluorescein angiography (FA), fundus photography (FP), and optical coherence tomography (OCT) from 2001 to 2009. Repeated-measures logistic regression was performed to compare patients' odds of undergoing these procedures in 2001, 2005, and 2009. In addition, the proportions of patients with an incident NVAMD or ME diagnosis in 2003 or 2008 who underwent FA, FP, and OCT were compared. Results. From 2001 to 2009, among patients with NVAMD, the odds of undergoing OCT increased 23-fold, whereas the odds of receiving FA and FP decreased by 68% and 79%, respectively. Similar trends were observed for ME. From 2003 to 2008, the proportion of patients undergoing OCT within 1 year of initial diagnosis increased by 315% for NVAMD and by 143% for ME; the proportion undergoing OCT without FA within 1 year increased by 463% for NVAMD and by 216% for ME. Conclusions. Use of OCT increased dramatically during the past decade, whereas use of FA and FP declined considerably, suggesting that OCT may be replacing more traditional diagnostic testing in patients with NVAMD or ME. Future studies should evaluate whether this increased reliance on OCT instead of FA and FP affects patient outcomes. PMID:24346174

Schneider, Eric W.; Mruthyunjaya, Prithvi; Talwar, Nidhi; Harris Nwanyanwu, Kristen; Nan, Bin; Stein, Joshua D.

2014-01-01

355

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

PubMed Central

An immunoglobulin G (IgG)–capture enzyme-linked immunosorbent assay (ELISA) for rubella virus is described. The assay uses a fluorescein isothiocyanate (FITC)–anti-FITC amplification system. The detection limit of the ELISA was approximately 7 IU of rubella virus-specific IgG per ml of serum sample. For saliva samples the performances of the capture ELISA and previously described radioimmunoassay were assessed, and the results of those two assays were compared to the rubella virus-specific IgG result obtained by a commercial ELISA (Behring Enzygnost) with a panel of paired serum and saliva samples. This comparison showed that the capture ELISA with saliva was more sensitive than the radioimmunoassay and that the results correlated better with the serum IgG result than the results of the radioimmunoassay did, with an overall sensitivity of 82% and a rank correlation of 0.68, whereas the sensitivity and rank correlation for the radioimmunoassay were 74% and 0.45, respectively. For subjects of 10 years of age or younger, the ELISA with saliva had a sensitivity of 94% and a specificity of 100% compared to the results of the ELISA (Behring Enzygnost) for rubella virus-specific IgG with corresponding serum samples. The sensitivity was much lower for subjects ages 17 years or older. The assay may have wider epidemiological use with saliva specimens, particularly those from children. PMID:9889225

Vyse, A. J.; Brown, D. W. G.; Cohen, B. J.; Samuel, R.; Nokes, D. J.

1999-01-01

356

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

NASA Astrophysics Data System (ADS)

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

Liu, Jia-ming; Lin, Li-ping; Jiang, Shu-Lian; Cui, Ma Lin; Jiao, Li; Zhang, Xiao Yang; Zhang, Li-hong; Zheng, Zhi Yong; Lin, Xuan; Lin, Shao-qin

2013-11-01

357

Fibrillin-1 staining anomalies are associated with increased staining for TGF-beta and elastic fibre degradation; new clues to the pathogenesis of emphysema.  

PubMed

We recently demonstrated aberrant staining of fibrillin-1 in lung tissue specimens with emphysematous lesions. In this study, we have extended this observation by an elaborate analysis of the elastic fibre. Using domain-specific antibodies to fibrillin-1, and to other elastin fibre-associated molecules, lung tissue derived from patients without obvious clinical emphysema, but harbouring various degrees of microscopical emphysematous lesions, was analysed. In addition, the fibrillin-regulated growth factor TGF-beta was studied. Electron microscopy and biochemical analysis of desmosine (a marker for elastin) were also performed. Results were compared with lung tissue derived from patients with clinical emphysema. Domain-specific antibodies recognizing the C-terminal, N-terminal, and middle part of fibrillin-1 showed aberrant staining patterns associated with increasing degrees of microscopical emphysema. Staining for elastin, emilin-1, and fibulin-2 was, however, not aberrant. TGF-beta staining was markedly increased. On the electron microscopic, but not light microscopical, level, initial elastic fibre degradation was noticed in specimens with microscopical emphysema. Lung specimens from patients with clinical emphysema also displayed fragmented fibrillin-1 staining and, in addition, displayed extensive degradation of the elastic fibre. The results suggest that fibrillin-1 anomalies and TGF-beta overexpression are associated with initial events occurring during the emphysematous process. Based on these and other data, a mechanism for emphysematogenesis is proposed. PMID:19373854

Koenders, Mieke M J F; Wismans, Ronnie G; Starcher, Barry; Hamel, Ben C J; Dekhuijzen, Richard P N; van Kuppevelt, Toin H

2009-08-01

358

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

PubMed

The staining mechanism of van Gieson's picrofuchsin was studied by use of simple protein model systems and tissue sections, and by spectrophotometry and dialysis experiments. At the endpoint of the staining reaction (equilibrium) cytoplasm is yellow. Dye dilution experiments demonstrated that the highest affinity in the tissue section--picrofuchsin system is between binding sites in cytoplasmic protein and acid fuchsin. Nevertheless sections that were first stained in acid fuchsin (AcF) and then in picrofuchsin ended up with cytoplasm stained yellow. It was concluded that differences in the dye diffusion rates and differences in the permeability of tissue components cannot be invoked to explain the differential staining result. Model experiments with dissolved proteins demonstrated a positive relationship between protein concentration and uptake of picric acid (PA) from picrofuchsin. From this and experiments with additives (sodium dodecylsulphate, urea etc.) and organic solvents, it is proposed that coagulant interchain cross-linking at the high protein concentration of the cytoplasm masks potential dye-binding sites. This affects high affinity dyes with multiple binding sites more than small dyes, and so puts AcF at a disadvantage compared to PA. Staining of non-collagen proteins is mainly by hydrophobic bonding, involving ionic attractions, apolar bonds, and release of water. This mode of binding is relatively strong, decreases swelling and leads to slow dye exchange. Dye binding to collagen is mostly by hydrogen bonds, but in aqueous dye solvent nonpolar residues and charged residues may also participate. This structure remains relatively open during and after dye-binding, and the bound dye ions are therefore easily exchanged for other dye ions. PMID:7683012

Prentø, P

1993-02-01

359

Cell-death assessment by fluorescent and nonfluorescent cytosolic and nuclear staining techniques.  

PubMed

Apoptosis, a genetically programmed cellular event leads to biochemical and morphological changes in cells. Alterations in DNA caused by several factors affect nucleus and ultimately the entire cell leading to compromised function of the organ and organism. DNA, a master regulator of the cellular events, is an important biomolecule with regards to cell growth, cell death, cell migration and cell differentiation. It is therefore imperative to develop the staining techniques that may lead to visualize the changes in nucleus where DNA is housed, to comprehend the cellular pathophysiology. Over the years a number of nuclear staining techniques such as propidium iodide, Hoechst-33342, 4', 6-diamidino-2-phenylindole (DAPI), Acridine orange-Ethidium bromide staining, among others have been developed to assess the changes in DNA. Some nonnuclear staining techniques such as Annexin-V staining, which although does not stain DNA, but helps to identify the events that result from DNA alteration and leads to initiation of apoptotic cell death. In this review, we have briefly discussed some of the most commonly used fluorescent and nonfluorescent staining techniques that identify apoptotic changes in cell, DNA and the nucleus. These techniques help in differentiating several cellular and nuclear phenotypes that result from DNA damage and have been identified as specific to necrosis or early and late apoptosis as well as scores of other nuclear deformities occurring inside the cells. PMID:24831993

Atale, N; Gupta, S; Yadav, U C S; Rani, V

2014-07-01

360

Staining potential of acidulated phosphate fluoride (APF) foam on dental restorations in vitro  

PubMed Central

Objectives: To identify the staining potential of acidulated phosphate fluoride (APF) foam on restorations in vitro. Materials and Methods: Two hundred ovine molars were used. Except 40 teeth remained unrestored as the controls, each was randomly selected to receive one of four restorative materials including preparation without restoration, glass ionomer cement (GIC), resin modified glass ionomer cement (RMGIC), or composite resin (CR). Following the procedure, topical APF was applied with a predetermined frequency. Staining formation was then evaluated. Results: APF-treated teeth and restorations appeared with a darker shade, an orange-colored surface and/or a brown margin. The staining rates on GIC, RMGIC, and CR were 50%, 27.5%, and 17.5%, respectively. GIC had a higher staining potential than RMGIC (?2 = 4.266, df = 1, P = 0.039) and CR (?2 = 9.448, df = 1, P = 0.002), whereas the difference between RMGIC and CR was indiscernible (?2 = 1.147, df = 1, P = 0.284). Repeated applications of topical APF increased the risk of staining on RMGIC (?2 = 8.436 df = 1, P = 0.004) and CR (?2 = 6.873, df = 1, P = 0.009) but not on GIC (?2 = 0, df = 1, P = 1) and the controls (?2 = 4.051, df = 3, P = 0.256). Conclusions: APF-foam-related staining was confirmed in vitro. GIC was more susceptible to fluoride staining. This study suggested aesthetic implications when applying fluorides to restored teeth. PMID:25657523

Lin, David; Huang, Boyen

2015-01-01

361

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

NASA Technical Reports Server (NTRS)

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

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

1991-01-01

362

The application of visible wavelength reflectance hyperspectral imaging for the detection and identification of blood stains.  

PubMed

Current methods of detection and identification of blood stains rely largely on visual examination followed by presumptive tests such as Kastle-Meyer, Leuco-malachite green or luminol. Although these tests are useful, they can produce false positives and can also have a negative impact on subsequent DNA tests. A novel application of visible wavelength reflectance hyperspectral imaging has been used for the detection and positive identification of blood stains in a non contact and non destructive manner on a range of coloured substrates. The identification of blood staining was based on the unique visible absorption spectrum of haemoglobin between 400 and 500nm. Images illustrating successful discrimination of blood stains from nine red substances are included. It has also been possible to distinguish between blood and approximately 40 other reddish stains. The technique was also successfully used to detect latent blood stains deposited on white filter paper at dilutions of up to 1 in 512 folds and on red tissue at dilutions of up to 1 in 32 folds. Finally, in a blind trial, the method successfully detected and identified a total of 9 blood stains on a red T-shirt. PMID:25498930

Li, Bo; Beveridge, Peter; O'Hare, William T; Islam, Meez

2014-12-01

363

Brilliant cresyl blue staining negatively affects mitochondrial functions in porcine oocytes.  

PubMed

Summary The aim of the present study was to examine the effects of brilliant cresyl blue (BCB) staining on mitochondrial functions in porcine oocytes. Cumulus-oocyte complexes (COCs) collected from slaughterhouse-derived porcine ovaries were cultured with (13 ?M) or without (0 ?M, control) BCB for 60 min. Mitochondrial functions in oocytes were examined immediately after staining or after in vitro maturation. The BCB-stained oocytes produced reactive oxygen species (ROS) at higher levels than control oocytes immediately after staining (2.2-fold, P < 0.001) and after maturation (1.7-fold, P < 0.001). The adenosine triphosphate (ATP) content and mitochondrial membrane potential (MMP) in oocytes were similar for the two groups immediately after staining. However, ATP and relative MMP levels were significantly (P < 0.05) lower in BCB-treated oocytes than in the control (2.18 versus 2.83 pM and 0.82 versus 1.0, respectively). There was no difference in mitochondrial DNA copy number between the two groups after maturation. The ATP content in early developmental stage embryos (3 days after parthenogenetic activation) was lower in the BCB-stained group than that in the control group but the difference was not significant. In conclusion, BCB staining of oocytes at the immature stage compromises mitochondrial functions throughout oocyte maturation, but function is restored during early embryo development. PMID:24355610

Santos, E C S; Sato, D; Lucia, T; Iwata, H

2013-12-20

364

Detecting glycogen in peripheral blood mononuclear cells with periodic Acid schiff staining.  

PubMed

Periodic acid Schiff (PAS) staining is an immunohistochemical technique used on muscle biopsies and as a diagnostic tool for blood samples. Polysaccharides such as glycogen, glycoproteins, and glycolipids stain bright magenta making it easy to enumerate positive and negative cells within the tissue. In muscle cells PAS staining is used to determine the glycogen content in different types of muscle cells, while in blood cell samples PAS staining has been explored as a diagnostic tool for a variety of conditions. Blood contains a proportion of white blood cells that belong to the immune system. The notion that cells of the immune system possess glycogen and use it as an energy source has not been widely explored. Here, we describe an adapted version of the PAS staining protocol that can be applied on peripheral blood mononuclear immune cells from human venous blood. Small cells with PAS-positive granules and larger cells with diffuse PAS staining were observed. Treatment of samples with amylase abrogates these patterns confirming the specificity of the stain. An alternate technique based on enzymatic digestion confirmed the presence and amount of glycogen in the samples. This protocol is useful for hematologists or immunologists studying polysaccharide content in blood-derived lymphocytes. PMID:25548935

Tabatabaei Shafiei, Mahdieh; Carvajal Gonczi, Catalina M; Rahman, Mohammed Samiur; East, Ashley; François, Jonathan; Darlington, Peter J

2014-01-01

365

The effectiveness of four methods for stain removal from direct resin-based composite restorative materials  

PubMed Central

Background/purpose Few studies investigated the best method for removing stains from different types of resin-based composite restorations and compared them to the more recently introduced nanocomposites. This study compared the effect of four methods for stain removal from composite resins; finishing with Sof-lex disks, using pumice and brush, bleaching with 10% carbamide peroxide and 38% hydrogen peroxide. Materials and methods Twenty disk specimens were prepared. Specimens were immersed in a staining solution for 3 weeks. The stained surfaces of five specimens from each RBC material were treated with one of the treatment procedures. Colorimetric measurements were taken using spectrophotometer prior to and after staining, and then repeated after surface treatments. Color difference values were calculated. Results One-way ANOVA indicated significant differences in color change of the three composite resin materials following staining. Filtek Z250 showed the least susceptibility to discoloration followed by Renamel, Filtek Supreme was the material most prone to discoloration. Two-way ANOVA and Tukey’s Post Hoc showed that all stain removing procedures except polishing with pumice, were able to return Filtek Z250 to clinically acceptable color difference. While bleaching with 38% carbamide peroxide was not effective with Renamel. Only pumice and 10% carbamide peroxide were able to return Renamel to clinically acceptable color difference. Conclusion Compositions of resin-based composite resins play an important role in their susceptibility to stain and their amenability to stain removal procedures. Home bleaching showed good results for the three materials, while office bleach was the least effective. PMID:24748758

Al-Nahedh, Hend Nahedh; Awliya, Wedad Yassin

2013-01-01

366

X-GAL STAINING [Adult Head Whole Mounts] Leslie Vosshall 12/21/2000  

E-print Network

M 50 uL 500 uL 5 mM K3Fe(CN)6 500 mM 50 uL 500 uL #12;3 5 mM K4Fe(CN)6-6H2O 500 mM 50 uL 500 uL X. Make up enough STAINING SOLUTION for 500 ul per tube. Add other components of staining solution and hold at 37 degrees until ready to incubate heads. · Add 500 ul STAINING SOLUTION per tube. Incubate

367

Staining procedure to aid in assessment of bucrylate histotoxicity in tissue sections.  

PubMed

Isobutyl-2-cyanoacrylate (bucrylate) is commonly used as a material for therapeutic embolization. Histologic sections of embolized tissue routinely stained with hematoxylin-eosin show the bucrylate as a translucent material. We outline a staining procedure that demonstrates bucrylate in tissue sections. It is useful in routinely prepared paraffin-embedded tissue, and it is also effective after embolized tissues have been processed with petroleum ether rather than xylene. This stain may be effectively utilized to assess harmful tissue reactions to bucrylate. PMID:2409951

Lundie, M J; Ellyatt, D; Kaufmann, J C; Vinters, H V

1985-08-01

368

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

SciTech Connect

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

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

2012-11-01

369

Evaluating the use of 3'-(p-Aminophenyl) fluorescein for determining the formation of highly reactive oxygen species in particle suspensions  

PubMed Central

Background Given the importance of highly reactive oxygen species (hROS) as reactants in a wide range of biological, photochemical, and environmental systems there is an interest in detection and quantification of these species. The extreme reactivity of the hROS, which includes hydroxyl radicals, presents an analytical challenge. 3'-(p-Aminophenyl) fluorescein (APF) is a relatively new probe used for measuring hROS. Here, we further evaluate the use of APF as a method for the detection of hydroxyl radicals in particle suspensions. Results Particle-generated hROS can be quantified with an estimated detection limit of 50 nM. Measurements of hROS in two National Institute of Standards and Technology (NIST 2709 and 2710) soil suspensions and a pyrite suspension show non-linear particle dose-response curves for hROS generation. APF can also be used in solutions containing no dissolved molecular oxygen (O2) to determine the role of O2 in the formation of hROS. Results confirm that O2 is mechanistically important in the formation of hROS by dissolved ferrous iron and in pyrite suspensions. Conclusion Given the non-linear dose-response curves for particle generation of hROS, we recommend using several particle loadings in experiments aimed to compare particles for their hROS generation potential. The method presented here is specific to hROS and simple to perform. The analysis can be conducted in mobile labs as only basic laboratory equipment is required. PMID:19671165

2009-01-01

370

High dose dietary pyridoxine induces T-helper type 1 polarization and decreases contact hypersensitivity response to fluorescein isothiocyanate in mice.  

PubMed

Pyridoxine (vitamin B(6)) is commonly used as a dietary supplement and beneficial effects of it are expected. However, excess ingestion of pyridoxine has been shown to cause a severe sensory neuropathy in humans and experimental animals. We have been studying the linkage between the nervous and immune systems using a fluorescein isothiocyanate (FITC)-induced contact hypersensitivity (CHS) mouse model. We have found that activation of transient receptor potential ankyrin 1 (TRPA1), which is expressed on sensory neurons, enhances skin sensitization to FITC. Another feature of FITC-induced CHS is its dependence on T helper 2 (Th2) type responses. We hypothesized that the excess intake of pyridoxine may affect sensitization to FITC and influence helper T-cell polarization. We examined FITC-induced CHS in BALB/c mice fed a diet containing excess pyridoxine (120 mg/kg diet) for 3 weeks. We found that mice fed on the excess-pyridoxine diet exhibited a lower response as to FITC-induced CHS compared with ones fed on a diet with a standard pyridoxine content (6.0 mg/kg diet). Moreover, the interferon (IFN)-?/interleukin (IL)-4 ratio produced by draining lymph node cells was significantly higher with the excess-pyridoxine diet. This suggested that the cytokine balance was shifted toward Th1 with the excess-pyridoxine diet. Consistently, Th1-dependent oxazolone-induced CHS was enhanced with the excess-pyridoxine diet. These results suggested that an excess pyridoxine intake actively influences the immune system by altering helper T cell polarization. PMID:22466557

Kobayashi, Chie; Kurohane, Kohta; Imai, Yasuyuki

2012-01-01

371

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

E-print Network

Targeted alteration of real and imaginary refractive index of biological cells by histological of epithe- lial cells caused by histological stains such as hematox- ylin and eosin-containing cytostain. We

Ottino, Julio M.

372

Limonite-stained Siltite near the Blackbird Cobalt-Copper Mine  

USGS Multimedia Gallery

Limonite-stained outcrop of the banded siltite unit of the Apple Creek Formation, near Blackbird Creek, south of the Blackbird cobalt-copper mine area, in the Salmon River Mountains of east-central Idaho....

373

What Poisoned the Apple Juice? A Gram Staining and Selective Media Lab.  

ERIC Educational Resources Information Center

Introduces an inquiry-based laboratory experiment in which students identify an unknown bacterial species by using techniques such as Gram staining. Uses an authentic problem solving approach in a scenario entitled, "What poisoned the apple juice?" (YDS)

Hammond, Paul; Brown, Nikole; Hauser, Doug; Pomart, Katrina; Karcher, Sue; Balschweid, Mark

2002-01-01

374

Rapid staining method to detect and identify downy mildew (Peronospora belbahrii) in basil1  

PubMed Central

• Premise of the study: Demand for fresh-market sweet basil continues to increase, but in 2009 a new pathogen emerged, threatening commercial field/greenhouse production and leading to high crop losses. This study describes a simple and effective staining method for rapid microscopic detection of basil downy mildew (Peronospora belbahrii) from leaves of basil (Ocimum basilicum). • Methods and Results: Fresh leaf sections infected with P. belbahrii were placed on a microscope slide, cleared with Visikol™, and stained with iodine solution followed by one drop of 70% sulfuric acid. Cell walls of the pathogen were stained with a distinct coloration, providing a high-contrast image between the pathogen and plant. • Conclusions: This new staining method can be used successfully to identify downy mildew in basil, which then can significantly reduce its spread if identified early, coupled with mitigation strategies. This technique can facilitate the control of the disease, without expensive and specialized equipment. PMID:25202569

Koroch, Adolfina R.; Villani, Thomas S.; Pyne, Robert M.; Simon, James E.

2013-01-01

375

Journal of Neuroscience Methods 153 (2006) 135146 A modified technique for high-resolution staining of myelin  

E-print Network

of colloidal silver to myelin for vi% formalin following impregnation in ammoniacal silver nitrate, and the use of a low concentration of 4 reveals similar patterns in the new myelin silver stain, the Gallyas stain, and myelin basic protein

Wang, Xiaoqin

376

Simplified heavy metal staining techniques demonstrated with Fast Plant leaf tissue  

Microsoft Academic Search

Fast Plant (Brassica rapa, Cruciferae) leaf tissue fixed in glutaraldehyde-acrolein and post-fixed in osmium, was examined for response to several easily-prepared heavy metal stains. Lead and uranium, separately and in combination, gave typical results across the spectrum of cell organelles. As a single stain following osmium, bismuth produced images seemingly equivalent to lead and uranium. Phosphotungstic acid produced very good

Joseph B. Harris; Thomas G. Guilliams; Jeffery A. Schultz

1992-01-01

377

Osmium tetroxide\\/p-phenylenediamine staining of nucleoli and balbiani rings in Chironomus salivary glands  

Microsoft Academic Search

This paper deals with the application of the osmium tetroxide fixation followed by p-phenylenediamine treatment to salivary gland cells from Chironomus larvae. After this procedure, cytoplasm, nucleoli and Balbiani rings show a high degree of staining both in light and electron microscopy, while chromatin remains unstained. Ethanol fixation followed by osmium tetroxide\\/p-phenylenediamine does not modify the above mentioned staining pattern.

Juan C. Stockert

1977-01-01

378

Evidence for wheat-rye nucleolar competition (amphiplasty) in triticale by silver-staining procedure  

Microsoft Academic Search

Amphiplasty in hexaploid triticale, the artificial amphiploid of tetraploid wheat and diploid rye, is analyzed for the first time using a modified, highly reproducible, silver-staining procedure. A comparative analysis of metaphase somatic cells by phase contrast, C-banding and silver-staining of the hexaploid triticale cv. ‘Cachirulo’ and its parents, namely, the tetraploid durum wheat cv. ‘Enano de Andujar’ and the diploid

J. R. Lacadena; M. C. Cermeño; J. Orellana; J. L. Santos

1984-01-01

379

Detection of alkali-silica reaction swelling in concrete by staining  

DOEpatents

A method using concentrated aqueous solutions of sodium cobalt nitrite and rhodamine B is described which can be used to identify concrete that contains gels formed by the alkali-silica reaction (ASR). These solutions present little health or environmental risk, are readily applied, and rapidly discriminate between two chemically distinct gels; K-rich, Na-K-Ca-Si gels are identified by yellow staining, and alkali-poor, Ca-Si gels are identified by pink staining.

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

1998-04-14

380

Detection of alkali-silica reaction swelling in concrete by staining  

DOEpatents

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

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

1998-01-01

381

Detection of single sequence repeat polymorphisms in denaturing polyacrylamide sequencing gels by silver staining  

Microsoft Academic Search

Large-scale use of molecular markers in plant breeding is limited by the throughput capacity for genotyping. DNA polymorphisms\\u000a can be detected in denaturing polyacrylamide gels indirectly by nucleotide labeling or directly by staining. Fluorescent-labeling\\u000a or radiolabeling requires sophisticated infrastructure not always available in developing countries. We present an improved\\u000a low-cost method for silver staining and compare it to 2 other

S. Creste; A. Tulmann Neto; A. Figueira

2001-01-01

382

Fluorescence changes during electrical activity in frog muscle stained with merocyanine  

Microsoft Academic Search

NERVE membranes stained with various fluorescent dyes show changes in fluorescence intensity concomitant with changes in transmembrane potential1-4. Cohen et al.4 described a number of dyes that exhibit changes in fluorescence emission which vary linearly with transmembrane potential. Two of these have been used to investigate the excitation-contraction process in skeletal muscle5-7. Whole muscles and single fibres stained with Nile

Julio Vergara; Francisco Bezanilla

1976-01-01

383

MicroCT of Coronary Stents: Staining Techniques for 3-D Pathological Analysis  

E-print Network

for the degree of MASTER OF SCIENCE May 2011 Major Subject: Biomedical Engineering MicroCT of Coronary Stents: Staining Techniques for 3-D Pathological Analysis Copyright 2011 Stephen Daniel Darrouzet... ABSTRACT MicroCT of Coronary Stents: Staining Techniques for 3-D Pathological Analysis. (May 2011) Stephen Daniel Darrouzet, B.S., Texas A&M Unversity Chair of Advisory Committee: Dr. Fred Clubb, Jr. In the area of translational research...

Darrouzet, Stephen 1987-

2011-04-21

384

MITOCHONDRIAL LOCALIZATION OF OXIDATIVE ENZYMES: STAINING RESULTS WITH TWO TETRAZOLIUM SALTS  

Microsoft Academic Search

A comparison is made of the staining results obtained with Nitro-BT and MTT-Co ++ as acceptors when DPNH is the substrate in frozen sections of cold formol-calcium-fixed rat kidney (normal and following ligation of the blood vessels) and human liver containing lipofuscin granules. The kidney results are evaluated in terms of mitoehondrial morphology seen after classical mitochondrial stains and in

ALEX B. NOVIKOFF; WOO-YUNG SHIN; JOAN DRUCKER

1961-01-01

385

Efficacy of LED versus KTP laser activation of photodynamic bleaching of tetracycline-stained dentine.  

PubMed

In some well-established laser applications where large spot sizes are used, an array of high-intensity light emitting diodes (LED) emitting at similar wavelength could potentially replace the laser. This situation applies for the photodynamic bleaching of stains in teeth. This study compared the relative efficacy of an array of visible green LED (535 nm?±?15 nm) with a KTP laser in photodynamic bleaching of tetracycline-stained dentine in human tooth roots. After establishing consistent staining in 96 roots using a validated method, the roots were sectioned into 2-3-mm thick horizontal slices that were treated with gels containing rhodamine B (Smartbleach® or Smartbleach® 3LT). Colour changes were tracked up to 1 month after treatment. While both systems were effective in bleaching the tetracycline-stained dentine, KTP laser activation gave greater bleaching efficacy than LED activation, enhancing the action of the gel. Use of the KTP laser would be preferable over an LED system when confronted with tetracycline staining. Use of this photodynamic bleaching method offers valuable means to reduce the severity of tetracycline staining. PMID:25288264

Bennett, Zackary Y; Walsh, Laurence J

2014-10-01

386

Glycol methacrylate embedding and microwave staining for light microscopy of the mouse cochlea.  

PubMed

This study examined the utility of a methacrylate-based embedding medium and microwave staining for light microscopic quantification of hair cells and spiral ganglion cells in the mouse cochlea. The most important phase of the preparation process involved slowing down the polymerization process. The tissue molecules so locked within the plastic matrix produced excellent preservation of the organ of Corti and adjacent structures including the spiral ganglion, as well as tissue ionic charges. Excitable by microwaves, these ionic charges accelerated the movement of the basic dye (hematoxylin) into the tissue, reducing the time for this segment of the staining process from approximately 45 minutes to 1-2 minutes. When embedded in glycol methacrylate (GMA), acidic dyes show less stain-cell affinity so that staining intensity and time cannot be improved significantly. However, addition of color extenders to the counterstain eosin produced distinguished staining of all tissue constituents. Thus, a combination of GMA embedding medium, use of the microwave for staining and addition of color extenders to the counterstain generated excellent structural resolution and contrast. This made both hair cell and spiral ganglion cell counts possible from within the same specimen and provided an opportunity for qualitative evaluation as well. PMID:7535474

Katbamna, B; Ralston, A

1994-01-01

387

X-gal staining of canine skin tissues: A technique with multiple possible applications  

PubMed Central

Background: Estimation of ?-galactosidase (?gal) activity in human cells and tissues indicate its possible use as a marker of senescence. Objectives: This study was done to detect senescence-associated ?gal (SA-?gal) activity in canine skin tissue by using its substrate 5-bromo-4-chloro-3-indolyl ?-D-galactosidase (X-gal). Materials and Methods: Skin samples were collected through rapid necropsy process. The X-gal staining was done by altering different factors of the staining procedure like pH of the reagents and incubation time. Further, effect of tissue preservation procedure was also evaluated. Results: Typical X-gal staining was detected in old dogs’ skin samples and it was detectable both at pH 6 and pH 7.3. The cells present in the inner lining of the hair follicles and sebaceous glands are the major cells that have high SA-?gal activity. The X-gal staining intensity was more prominent in tissues preserved in liquid nitrogen at -196°C than in -80°C freezer. Prolonged incubation period increased the intensity of staining. Conclusions: This study indicates possibility of X-gal staining in canine tissues and opens an avenue for further in-depth studies that might be useful for different research and clinical studies like determination of dog's approximate age. PMID:25097391

Pati, Soumyaranjan; Jain, Sumeet; Behera, Monalisa; Acharya, Aditya Prasad; Panda, Susen K.; Senapati, Shantibhusan

2014-01-01

388

Lipid content, active mitochondria and brilliant cresyl blue staining in bovine oocytes.  

PubMed

Bovine oocytes that stain with brilliant cresyl blue (BCB) have a relatively higher developmental competence. The aim of the present study was to investigate the relationships among BCB staining, lipid content, and active mitochondria. Bovine oocytes (N = 133) with at least three layers of cumulus cells were segregated as BCB retained (BCB+) or metabolized (BCB-) and then stained for active mitochondria (Mitotracker Red) and lipid (Bodipy), with analysis by confocal microscopy. The BCB+ oocytes (N = 45) contained approximately 26% more cytoplasmic lipid than BCB- oocytes (N = 26-27; P < 0.05). Staining for active mitochondria did not differ between the groups. In BCB- oocytes but not BCB+ oocytes, lipid content correlated with active mitochondrial staining (r = 0.48; P < 0.05). Diameter correlated with lipid content for BCB+ oocytes (r = 0.46; P < 0.05), but not for BCB- oocytes (r = 0.16; P > 0.05). Irrespective of BCB staining, both lipid and active mitochondrial content correlated with diameter. In conclusion, the higher lipid content of BCB+ bovine oocytes might provide a cellular and functional basis for their greater developmental competence. PMID:23199746

Castaneda, Cesar A; Kaye, Peter; Pantaleon, Marie; Phillips, Nancy; Norman, Scott; Fry, Richard; D'Occhio, Michael J

2013-02-01

389

Sweet silver: a formaldehyde-free silver staining using aldoses as developing agents, with enhanced compatibility with mass spectrometry.  

E-print Network

Sweet silver: a formaldehyde-free silver staining using aldoses as developing agents, with enhanced are also favored features. Silver staining combines many of these features, but its compatibility with mass spectrometry is limited. We describe here a new variant of silver staining that is completely formaldehyde

Paris-Sud XI, Université de

390

The application of chemical staining to separate calcite and aragonite minerals for micro-scale isotopic analyses  

Microsoft Academic Search

Staining is a useful laboratory tool for distinguishing specific carbonate minerals from a mixture of carbonates. Feigl's and Meigen's solutions are commonly used to distinguish aragonite from other carbon- ates. Feigl's solution stains the aragonite surface black and Meigen's stains purple, whereas the coexisting calcite remains nochang in color with either solutions. Both methods were applied to an authigenically precipitated

KAZUHIRO KATO; HIDEKI WADA; KANTARO FUJIOKA

391

Fibre sizes and histochemical staining characteristics in normal and chronically stimulated fast muscle of cat.  

PubMed Central

1. Normal and chronically stimulated peroneus longus muscles of the cat's hind limb were studied with respect to fibre size and staining properties for myofibrillar (myosin) adenosine triphosphatase (ATPase) and succinate dehydrogenase (SDH) activity. The intensity of staining for SDH activity was measured by microphotometry from the central portions of the muscle fibres ('core-SDH staining'). For comparison, histochemical properties were also studied in non-stimulated soleus muscles. 2. On account of the pH sensitivity of their myofibrillar ATPase, about 18% of the fibres in normal peroneus longus muscles were classified as type I, and about half of the remainder as II A and II B respectively. 3. In the normal peroneus longus muscles, the mean diameter of single muscle fibres generally varied between about 25 and 75 micron, whereby the average size of type I less than type II. 4. In the normal peroneus longus muscles the staining intensity for core SDH varied over a wide range. The average heaviness of staining was clearly ranked in the order type I greater than type II A greater than type II B. 5. Chronic stimulation was given to the deafferented common peroneal nerve by aid of a portable and remotely controlled mini-stimulator. The stimulation was delivered in 'tonic' patterns (greater than or equal to 50% of total time taken up by activity) of 'fast' (20 or 40 Hz) or 'slow' (5 or 10 Hz) rates. 6. Prior to the period of long-term stimulation, the cats had been subjected to a dorsal rhizotomy and hemispinalization on the ipsilateral (left) side. In the absence of chronic stimulation, these operations had no evident effects on the sizes or staining properties of peroneus longus fibres. 7. After 8 weeks of treatment with tonic patterns of stimulation, the fibres of peroneus longus muscles clearly became more similar to each other with respect to their diameter as well as their staining for ATPase and SDH activity. With respect to ATPase staining, however, the chronically stimulated peroneus longus fibres had become more similar to non-stimulated soleus fibres than to non-stimulated type I fibres of peroneus longus. With respect to the staining for core SDH, the chronically stimulated fibres all became similar to normal II A fibres of peroneus longus. The 'fast' and 'slow' patterns of chronic stimulation had the same effects on the staining properties. 8. Chronically stimulated peroneus longus muscles showed a decrease in fibre diameter which corresponded, roughly, to the concomitant decrease in muscle weight.(ABSTRACT TRUNCATED AT 400 WORDS) Images Fig. 1 Fig. 3 Fig. 5 PMID:2957493

Donselaar, Y; Eerbeek, O; Kernell, D; Verhey, B A

1987-01-01

392

Iodine Vapor Staining for Atomic Number Contrast in Backscattered Electron and X-ray Imaging  

PubMed Central

Iodine imparts strong contrast to objects imaged with electrons and X-rays due to its high atomic number (53), and is widely used in liquid form as a microscopic stain and clinical contrast agent. We have developed a simple technique which exploits elemental iodine's sublimation-deposition state-change equilibrium to vapor stain specimens with iodine gas. Specimens are enclosed in a gas-tight container along with a small mass of solid I2. The bottle is left at ambient laboratory conditions while staining proceeds until empirically determined completion (typically days to weeks). We demonstrate the utility of iodine vapor staining by applying it to resin-embedded tissue blocks and whole locusts and imaging them with backscattered electron scanning electron microscopy (BSE SEM) or X-ray microtomography (XMT). Contrast is comparable to that achieved with liquid staining but without the consequent tissue shrinkage, stain pooling, or uneven coverage artefacts associated with immersing the specimen in iodine solutions. Unmineralized tissue histology can be read in BSE SEM images with good discrimination between tissue components. Organs within the locust head are readily distinguished in XMT images with particularly useful contrast in the chitin exoskeleton, muscle and nerves. Here, we have used iodine vapor staining for two imaging modalities in frequent use in our laboratories and on the specimen types with which we work. It is likely to be equally convenient for a wide range of specimens, and for other modalities which generate contrast from electron- and photon-sample interactions, such as transmission electron microscopy and light microscopy. Microsc. Res. Tech. 77:1044–1051, 2014. © 2014 The Authors. Microscopy Research Technique published by Wiley Periodocals, Inc. PMID:25219801

Boyde, Alan; Mccorkell, Fergus A; Taylor, Graham K; Bomphrey, Richard J; Doube, Michael

2014-01-01

393

Dual AO/EB Staining to Detect Apoptosis in Osteosarcoma Cells Compared with Flow Cytometry  

PubMed Central

Background The aim of this study was to evaluate the ability of dual acridine orange/ethidium bromide (AO/EB) staining to detect tumor cell apoptosis. According to apoptosis-associated changes of cell membranes during the process of apoptosis, a clear distinction is made between normal cells, early and late apoptotic cells, and necrotic cells. Material/Method We cultured human osteosarcoma cells with 30, 60, and 120 ?g/ml kappa-selenocarrageenan. To assess the rates of cell proliferation and apoptosis, cells were fluorescently stained with acridine orange/ethidium bromide (AO/EB) or stained with propidium iodide (PI) and analyzed by flow cytometry. All experiments were repeated at least 3 times. Result Normal tumor cells, early and late apoptotic cells, and necrotic cells were examined using fluorescent microscopy. Early-stage apoptotic cells were marked by crescent-shaped or granular yellow-green acridine orange nuclear staining. Late-stage apoptotic cells were marked with concentrated and asymmetrically localized orange nuclear ethidium bromide staining. Necrotic cells increased in volume and showed uneven orange-red fluorescence at their periphery. Cells appeared to be in the process of disintegrating. The percentage of apoptotic osteosarcoma cells detected by dual acridine orange/ethidium bromide (AO/EB) staining was not significantly different from that detected using flow cytometry (P>0.05). Conclusions Our results suggest that dual acridine orange/ethidium bromide staining is an economic and convenient method to detect apoptosis in tumor cells and to test tumor chemosensitivity compared with flow cytometry. PMID:25664686

Liu, Kuan; Liu, Peng-cheng; Liu, Run; Wu, Xing

2015-01-01

394

Iodine vapor staining for atomic number contrast in backscattered electron and X-ray imaging.  

PubMed

Iodine imparts strong contrast to objects imaged with electrons and X-rays due to its high atomic number (53), and is widely used in liquid form as a microscopic stain and clinical contrast agent. We have developed a simple technique which exploits elemental iodine's sublimation-deposition state-change equilibrium to vapor stain specimens with iodine gas. Specimens are enclosed in a gas-tight container along with a small mass of solid I2 . The bottle is left at ambient laboratory conditions while staining proceeds until empirically determined completion (typically days to weeks). We demonstrate the utility of iodine vapor staining by applying it to resin-embedded tissue blocks and whole locusts and imaging them with backscattered electron scanning electron microscopy (BSE SEM) or X-ray microtomography (XMT). Contrast is comparable to that achieved with liquid staining but without the consequent tissue shrinkage, stain pooling, or uneven coverage artefacts associated with immersing the specimen in iodine solutions. Unmineralized tissue histology can be read in BSE SEM images with good discrimination between tissue components. Organs within the locust head are readily distinguished in XMT images with particularly useful contrast in the chitin exoskeleton, muscle and nerves. Here, we have used iodine vapor staining for two imaging modalities in frequent use in our laboratories and on the specimen types with which we work. It is likely to be equally convenient for a wide range of specimens, and for other modalities which generate contrast from electron- and photon-sample interactions, such as transmission electron microscopy and light microscopy. PMID:25219801

Boyde, Alan; Mccorkell, Fergus A; Taylor, Graham K; Bomphrey, Richard J; Doube, Michael

2014-12-01

395

[Evaluation of Hucker stain as Campylobacter sp screening detection during an acute diarrhea disease].  

PubMed

Campylobacter infection is one of the most frequent causes of gastroenteritis in the world and the third in Chile according to some studies. The routinary culture for Campylobacter in our country is not performed because of its high cost, and it is known, that the Hucker stain is a reasonable screening alternative. The objective of this study was to know the utility of the Hucker stain and estimate the frequency of Campylobacter in stool samples. A total of 5,750 stool samples received in the Catholic University Health Net Microbiology Laboratories, from March 2002 to May 2004, were studied with conventional stool culture and Hucker stain. In order to validate the Hucker stain with culture, during one month, all the stool samples were also studied with Campylobacter culture, with 35% sensitivity and 100% specificity. In 115/5.750 samples (2%), curved bacilli suggesting Campylobacter were observed by Hucker stain, and another 233 enteropathogens (4%) which corresponded to: 151 Salmonella sp, 55 Shigella sp, 25 enterohemorrhagic E coli, and 2 Yersinia sp were isolated. When analyzing the patients in whom the Hucker stain was positive, 62.2% were younger than 5 years and of these, 63.8% were infants. We conclude that the Hucker stain is a simple and specific method, although not very sensitive, that allows us to increase the yield of diagnosing enteric pathogens in a 33%. Campylobacter sp was in the second place after Salmonella sp in stool samples, and most frequent in young children. The active search for Campylobacter by means of culture is fundamental in the diagnosis of acute diarrhea. PMID:16077891

Chanqueo C, Leonardo; García C, Patricia; León C, Eugenia; Blu F, Antonieta

2005-09-01

396

Diagnosis of bacterial vaginosis by direct gram stain of vaginal fluid.  

PubMed

To determine whether bacterial vaginosis (BV), also known as nonspecific vaginitis, could be diagnosed by evaluating a Gram stain of vaginal fluid, we examined samples from 60 women of whom 25 had clinical evidence of BV and 35 had candidal vaginitis or normal examinations. An inverse relationship between the quantity of the Lactobacillus morphotype (large gram-positive rods) and of the Gardnerella morphotype (small gram-variable rods) was noted on Gram stain (P less than 0.001). When Gram stain showed a predominance (3 to 4+) of the Lactobacillus morphotype with or without the Gardnerella morphotype, it was interpreted as normal. When Gram stain showed mixed flora consisting of gram-positive, gram-negative, or gram-variable bacteria and the Lactobacillus morphotype was decreased or absent (0 to 2+), the Gram stain was interpreted as consistent with BV. Gram stain was consistent with BV in 25 of 25 women given a clinical diagnosis of BV and in none of 35 women with candidal vaginitis or normal examinations. Duplicate slides prepared from 20 additional specimens of vaginal fluid were stained by two methods and examined by three evaluators. Interevaluator interpretations and intraevaluator interpretations of duplicate slides were in agreement with one another and with the clinical diagnosis greater than or equal to 90% of the time. We concluded that a microscopically detectable change in vaginal microflora from the Lactobacillus morphotype, with or without the Gardnerella morphotype (normal), to a mixed flora with few or no Lactobacillus morphotypes (BV) can be used in the diagnosis of BV. PMID:6193137

Spiegel, C A; Amsel, R; Holmes, K K

1983-07-01

397

Oxidized p-phenylenediamine: observations on the staining reaction in epoxy embedded tissues.  

PubMed

p-Phenylenediamine (PPD) is easily oxidized to brown compounds which stain acidic substrates. On account of the spontaneous oxidation process, the colour of PPD increases and becomes ochre-brown in a few days, showing an absorption peak at lambda = 510 nm with shoulder at about 440 to 460 nm. Studies on the application of oxidized PPD as a stain for semi-thin sections revealed that some tissue components could be clearly visualized. After glutaraldehyde fixation, semi-thin and thin sections of animal tissues were treated with 0.5% aqueous PPD solutions which were aged for variable times at room temperature. Microvilli, goblet cell mucin, mast cell granules, cartilage matrix, collagen, elastin, keratohyalin granules, acrosomes, cytoplasmic granules of Drosophila hydei salivary glands and chromatin showed positive staining reactions after treatment of semi-thin sections with oxidized PPD (7-10 days aged) for 20-30 minutes. Microspectrophotometric studies revealed an absorption peak at lambda = 520-530 nm and a shoulder at lambda = 440-460 nm in goblet cell mucin stained by oxidized PPD. In the presence of anionic macromolecules, the main peak of oxidized PPD solutions showed a strong hyperchromism. Thin sections stained by oxidized PPD did not appear contrasted, but the treatment with 0.125% gold chloride (AuCl3) induced massive gold deposits in structures stained by oxidized PPD. Hyperchromic shifts were also produced in oxidized PPD solutions after the addition of small amounts of AuCl3. This procedure can be used as a simple and rapid staining method for epoxy sections, giving selective contrast for some tissue components. PMID:2422692

Armas-Portela, R; Stockert, J C; Ferrer, J M; Tato, A; Gómez-Perretta, C; Callaghan, R C

1986-01-01

398

Use of DNA stains in immunophenotyping by slide-based cytometry  

NASA Astrophysics Data System (ADS)

Immunophenotyping of peripheral blood leukocytes (PBL) is a very well documented application of Slide Based Cytometry (SBC). As for any other assay it is of highest importance to ensure that all cells which are relevant for an analysis are recognized. Unlike assays for cultured cells which have homogenous morphology immunophenotyping of PBLs is performed on cells with heterogeneous size and shape. Therefore, triggering on parameters related to cell morphology might lead to an incomplete analysis of just a subset of cells especially in pathological conditions. Several dyes stain DNA specifically in a wide variety of emission spectra. Many of them show some influence of the chromatin condensation and organization on the staining intensity. DNA dyes therefore can be used to differentiate between cell types having the same ploidy. This can be exploited for immunophenotyping since some dyes therefore can partially replace antibody staining. The concept of using DNA dyes in the setting of immunostaining has the following advantages: (1) nuclear staining provides a stable and easy triggering signal that guarantees both, that neither cells are excluded nor that debris or polluting particles are included into the analysis; (2) some DNA dyes differentiate between mononuclear and polymorphonuclear cells. A disadvantage of DNA dyes is that mostly cells have to be permeabilized. Because of this only one set of immunophenotypic markers can be stained, cells are fixed and permeabilized, and then nuclei are stained with the appropriate DNA dye. In the study we demonstrate the use of the most commonly available DNA dyes (7-AAD, To-Pro, To-To, PI etc.) in their applicability in immunophenotyping. An overview of spectral properties, fluorescence spill-over and optimal combinations with surface antigen staining will be shown. As in general for SBC only very small sample volumes are needed. This allows to serially analyze PBL in clinical settings that up to now could not be studied in detail such as in the critical ill patient, during major surgery, and in new-borns and infants.

Gerstner, Andreas O. H.; Laffers, Wiebke; Bootz, Friedrich; Tarnok, Attila

2003-06-01

399

A rapid stain-clearing method for video based cytological analysis of cotton megagametophytes.  

PubMed

Optical "clearing" is a cost saving method for preparing large numbers of whole, dissected or thickly sectioned cytological specimens such as plant ovules and ovaries. Minimal labor is required and specimens retain three-dimensional integrity. Previous development of high contrast stain-clearing methods using hemalum to impart contrast has facilitated analysis and photography under bright field illumination for small ovules. The deep stain intensity of hemalum, however, often precludes adequate light transmission and contrast within internal focal planes, limiting the applicability of hemalum-based stain-clearing to small specimens. Having encountered this problem for nucelli of cotton (Gossypium barbadense L.), which are roughly 300 microns thick at fertilization, we have developed a modified stain-clearing system. The two key features of these new methods are the use of azure, C, which allows the intensity of staining to be readily regulated, and contrast manipulation via video signal and image processing. Intensity of azure C stain was readily controlled by modifying the staining and/or dehydration media to produce relatively low contrast specimens. Analysis was facilitated by indirect viewing on a video monitor using adjustments of sensitivity, exposure, and contrast of the charge-coupled device (CCD) camera. Digital processing provided further enhancement. Acceptable images were obtained from virtually all specimens. These methods, which combine low contrast (high transmittance) specimens with high contrast imaging, should facilitate data acquisition on reproduction, thus the developmental and genetic characterization of reproductive mutants. Other applications, e.g., in pathology and embryology, are readily envisioned. PMID:9062705

Hodnett, G L; Crane, C F; Stelly, D M

1997-01-01

400

A simple, rapid and low-cost staining method for gel-electrophoresis separated phosphoproteins via the fluorescent purpurin dye.  

PubMed

A novel fluorescence detection method for phosphoproteins in 1-D and 2-D SDS-PAGE by using purpurin is developed in this study. Phosphoproteins as low as 4-8 ng could be specifically detected by purpurin within 60 min, and the detection limit is similar to or better than that of Pro-Q Diamond staining. Only 2 steps (staining and destaining) are needed for purpurin staining without requiring excessive fixing and washing steps, and for single use, $0.8 is enough for purpurin staining. By comprehensively comparing with Pro-Q Diamond staining, it is concluded that purpurin staining is a simple, rapid and low-cost staining method for a broad application to the research of phosphoproteins. PMID:25325196

Cong, Weitao; Shen, Jiayi; Xuan, Yuanhu; Zhu, Xinliang; Ni, Maowei; Zhu, Zhongxin; Hong, Guoying; Lu, Xianghong; Jin, Litai

2014-12-01

401

Quantitative assessment of relative changes of immunohistochemical staining by light microscopy in specified anatomical regions.  

PubMed

Despite the advent of ever newer microscopic techniques for the study of the distribution of macromolecules in biological tissues, the enzyme-based immunohistochemical (IHC) methods are still used widely and routinely. However, the acquisition of reliable conclusions from the pattern of the reaction products of IHC procedures is hindered by the regular need for subjective judgments, in view of frequent inconsistencies in staining intensity from section to section or in repeated experiments. Consequently, when numerical comparisons are required, light microscopic morphological descriptions are commonly supplemented with analytical data (e.g. from Western blot analyses); however, these cannot be directly associated with accurate structural information and can easily be contaminated with data from outside the region of interest. Alternatively, to eliminate the more or less subjective evaluation of the results of IHC staining, procedures should be developed that correct for the variability of staining through the use of objective criteria. This paper describes a simple procedure, based on digital image analysis methods and the use of an internal reference area on the analyzed sections, that reduces the operator input and hence subjectivity, and makes the relative changes in IHC staining intensity in different experiments comparable. The reference area is situated at a position of the section that is not affected by the experimental treatment, or a disease condition, and that can therefore be used to specify the baseline of the IHC staining. Another source of staining variability is the internal heterogeneity of the object to be characterized, which means that identical fields can never be analyzed. To compensate for this variability, details are given of a systematic random sampling paradigm, which provides simple numerical data describing the extent and strength of IHC staining throughout the entire volume to be characterized. In this integrated approach, the figures are derived by pooling relative IHC staining intensities from all sections of the series from a particular animal. The procedure (1) eliminates the problem arising from the personal assessment of the significance of the IHC staining intensity, (2) does not depend on the precise dissection of the tissue on a gross scale and (3) considerably reduces the consequences of limited, arbitrary sampling of the region of interest for IHC analysis. The quantification procedure is illustrated by data from an experiment in which inflammatory reactions in the murine spinal cord, measured as microglial activation, were followed by IHC after the lesion of the sciatic nerve. PMID:19335461

Paizs, M; Engelhardt, J I; Siklós, L

2009-04-01

402

New versatile staining reagents for biological transmission electron microscopy that substitute for uranyl acetate.  

PubMed

Aqueous uranyl acetate has been extensively used as a superb staining reagent for transmission electron microscopy of biological materials. However, recent regulation of nuclear fuel material severely restricts its use even for purely scientific purposes. Since uranyl salts are hazardous due to biological toxicity and remaining radioactivity, development of safe and non-radioactive substitutes is greatly anticipated. We examined two lanthanide salts, samarium triacetate and gadolinium triacetate, and found that 1-10% solution of these reagents was safe but still possess excellent capability for staining thin sections of plastic-embedded materials of animal and plant origin. Although post-fixation with osmium tetroxide was essential for high-contrast staining, post-staining with lead citrate could be eliminated if a slow-scan CCD camera is available for observation. These lanthanide salts can also be utilized as good negative-staining reagents to study supramolecular architecture of biological macromolecules. They were not as effective as a fixative of protein assembly, reflecting the non-hazardous nature of the reagents. PMID:22146677

Nakakoshi, Masamichi; Nishioka, Hideo; Katayama, Eisaku

2011-12-01

403

Early colonic dysplasia: comparison of differential mucin staining and tritiated thymidine labeling  

SciTech Connect

Controversy has arisen regarding the interpretation and significance of histochemical changes in the mucin produced by the globlet cells in colonic mucosa. The shift from sulfomucin to sialomucin, which is readily identified utilizing high iron diamine-alcian blue staining techniques, has been alternately interpreted as a specific, early dysplastic and premalignant change or a nonspecific generalized response to trauma and inflammation, among others. An attempt to clarify this issue was made by comparing mucin changes identified by high iron diamine-alcian blue staining techniques with increases in DNA synthetic activity identified utilizing autoradiographic analysis of tritiated thymidine uptake. Male Holtzman rats were treated with 15 weekly subcutaneous injections of dimethylhydrazine (30 mg/kg per week) (10 rats) or placebo (10 rats). The colons were prepared and fixed, sequential sections were stained with hematoxylin-eosin or high iron diamine-alcian blue, autoradiography was performed. Analyses of labeling index showed no difference in normal background crypts between the control and treatment groups nor in crypts adjacent to those displaying abnormal mucin staining. Crypts with abnormal mucin production (sialomucin dominant) had significantly higher labeling indexes when compared with those of control animals (p less than 0.005). These findings indicate that the shifts in mucin production identified with high iron diamine-alcian blue staining represent crypts with increased and abnormally distributed mitotic activity that is an early dysplastic response to the carcinogenic stimulus.

Chabot, J.A.; Colacchio, T.A.

1985-01-01

404

Optimal Iodine Staining of Cardiac Tissue for X-Ray Computed Tomography  

PubMed Central

X-ray computed tomography (XCT) has been shown to be an effective imaging technique for a variety of materials. Due to the relatively low differential attenuation of X-rays in biological tissue, a high density contrast agent is often required to obtain optimal contrast. The contrast agent, iodine potassium iodide (), has been used in several biological studies to augment the use of XCT scanning. Recently was used in XCT scans of animal hearts to study cardiac structure and to generate 3D anatomical computer models. However, to date there has been no thorough study into the optimal use of as a contrast agent in cardiac muscle with respect to the staining times required, which has been shown to impact significantly upon the quality of results. In this study we address this issue by systematically scanning samples at various stages of the staining process. To achieve this, mouse hearts were stained for up to 58 hours and scanned at regular intervals of 6–7 hours throughout this process. Optimal staining was found to depend upon the thickness of the tissue; a simple empirical exponential relationship was derived to allow calculation of the required staining time for cardiac samples of an arbitrary size. PMID:25170844

Zhang, Yanmin; Lei, Ming; Withers, Philip J.; Zhang, Henggui

2014-01-01

405

Use of urinary gram stain for detection of urinary tract infection in childhood.  

PubMed Central

In this study, urinary culture, urinary Gram stain, and four tests within the urinalysis, leukocyte esterase, nitrite, microscopyfor bacteria, and microscopyforpyuria, were examined in 100 children with symptoms suggesting urinary tract infection. Our purpose was to determine the validity of the urinary Gram stain compared with a combination of pyuria plus Gram stain and overall urinalysis (positiveness of nitrite, leukocyte esterase, microscopy for bacteria, or microscopy for white blood cell). Of 100 children, aged two days to 15 years, 70 (70 percent) had a positive urinary culture: 40 girls (57 percent) and 30 boys (43 percent). Escherichia coli was the most common isolated agent. The sensitivity and specificity of the urinary Gram stain were 80 percent and 83 percent, and that of the combination of pyuria plus Gram stain 42 percent and 90 percent, and that of the overall urinalysis 74 percent and 3.5 percent respectively. Our findings revealed that neither method of urine screen should substitute for a urine culture in the symptomatic patients in childhood. PMID:12230312

Arslan, Sükrü; Caksen, Hüseyin; Rastgeldi, Levent; Uner, Abdurrahman; Oner, Ahmet Faik; Odaba?, Dursun

2002-01-01

406

Gram stain as a relapse predictor of bacterial vaginosis after metronidazole treatment.  

PubMed

Bacterial vaginosis is the most prevalent disease of the female genital tract. In spite of various effective antibiotics in the treatment of bacterial vaginosis, its high relapse rate is a common problem. Bacterial species causing bacterial vaginosis are generally unable to be cultured by conventional methods. It is impractical and inadequate to use culture methods to guide initial treatment. Gram stain of vaginal secretion is a practical tool to establish the diagnosis of bacterial vaginosis. We enrolled 78 cases of Gram stain-proven bacterial vaginosis and tried to use Gram stain as a predictor of relapse after 1 week of treatment with metronidazole. Possible predictive factors for relapse in Gram stain were analyzed, including absence of large Gram-positive rods, presence of small Gram-negative rods, small Gram-variable rods, curved Gram-variable rods, or Gram-positive cocci. Gram stain was repeated immediately after treatment, and at 1 month and 3 months after treatment. All cases showed beneficial clinical effect after metronidazole treatment. Eighteen cases (23.1%) relapsed during the follow-up period. All 16 cases with significant Gram-positive cocci in pretreatment smears relapsed after metronidazole treatment. Presence of small Gram-negative rods, small Gram-variable rods, and curved Gram-variable rods, or absence of large Gram-positive rods did not predict relapse. Gram-positive cocci in pretreatment smear was a good predictor of relapse after metronidazole treatment. PMID:15843859

Huang, Mei; Wang, Jen Hsien

2005-04-01

407

Optimal iodine staining of cardiac tissue for X-ray computed tomography.  

PubMed

X-ray computed tomography (XCT) has been shown to be an effective imaging technique for a variety of materials. Due to the relatively low differential attenuation of X-rays in biological tissue, a high density contrast agent is often required to obtain optimal contrast. The contrast agent, iodine potassium iodide ([Formula: see text]), has been used in several biological studies to augment the use of XCT scanning. Recently I2KI was used in XCT scans of animal hearts to study cardiac structure and to generate 3D anatomical computer models. However, to date there has been no thorough study into the optimal use of I2KI as a contrast agent in cardiac muscle with respect to the staining times required, which has been shown to impact significantly upon the quality of results. In this study we address this issue by systematically scanning samples at various stages of the staining process. To achieve this, mouse hearts were stained for up to 58 hours and scanned at regular intervals of 6-7 hours throughout this process. Optimal staining was found to depend upon the thickness of the tissue; a simple empirical exponential relationship was derived to allow calculation of the required staining time for cardiac samples of an arbitrary size. PMID:25170844

Butters, Timothy D; Castro, Simon J; Lowe, Tristan; Zhang, Yanmin; Lei, Ming; Withers, Philip J; Zhang, Henggui

2014-01-01

408

Specific in vivo staining of astrocytes in the whole brain after intravenous injection of sulforhodamine dyes.  

PubMed

Fluorescent staining of astrocytes without damaging or interfering with normal brain functions is essential for intravital microscopy studies. Current methods involved either transgenic mice or local intracerebral injection of sulforhodamine 101. Transgenic rat models rarely exist, and in mice, a backcross with GFAP transgenic mice may be difficult. Local injections of fluorescent dyes are invasive. Here, we propose a non-invasive, specific and ubiquitous method to stain astrocytes in vivo. This method is based on iv injection of sulforhodamine dyes and is applicable on rats and mice from postnatal age to adulthood. The astrocytes staining obtained after iv injection was maintained for nearly half a day and showed no adverse reaction on astrocytic calcium signals or electroencephalographic recordings in vivo. The high contrast of the staining facilitates the image processing and allows to quantify 3D morphological parameters of the astrocytes and to characterize their network. Our method may become a reference for in vivo staining of the whole astrocytes population in animal models of neurological disorders. PMID:22509398

Appaix, Florence; Girod, Sabine; Boisseau, Sylvie; Römer, Johannes; Vial, Jean-Claude; Albrieux, Mireille; Maurin, Mathieu; Depaulis, Antoine; Guillemain, Isabelle; van der Sanden, Boudewijn

2012-01-01

409

Evaluation of agonal artifacts in the myocardium using a combination of histological stains and immunohistochemistry.  

PubMed

The problem of discrimination between agonal artifacts and intravital ischemic myocardial lesions was studied with four histochemical stains [hematoxylin-eosin, Mallory's phosphotungstic acid hematoxylin (PTAH), modified luxol fast blue, and Lie's hematoxylin basic fuchsin picric acid (HBFP)] and with immunohistochemistry using two antibodies (antimyoglobin and anti-C5b-9). Seventy-five forensic autopsy cases were divided into six groups designed to represent successively shorter periods of agonal myocardial ischemia: (a) sudden deaths with coronary artery disease, macroscopically visible myocardial infarction, and/or fresh coronary thrombus; (b) unexplained sudden deaths without coronary artery disease; (c) accidental CO poisoning; (d) suicidal CO poisoning in cars; (e) suicidal hangings; and (f) instant traumatic deaths, i.e., total brainstem laceration or rupture of the thoracic aorta. From each heart, five pieces were removed from standardized locations, and six parallel sections were stained with each method (i.e., 30 sections from each heart). Hematoxylin-eosin and anti-C5b-9 were only positive in the first three groups, thus indicating specificity for intravital necrotic changes. The other staining methods were "positive" in one or more cases in all six groups, thus implicating a high degree of sensitivity for artifactual, agonal ischemic changes. The latter methods cannot be used alone in the diagnosis of myocardial infarction. By staining parallel sections with different stains and antibodies, it seems possible to estimate the relative length of the agonal period in cardiac and noncardiac deaths. PMID:9185934

Edston, E

1997-06-01

410

PHOSPHOLIPID AND FDA ACTIVITY MEASUREMENTS ADAPTED TO BIOLOGICAL GAC  

EPA Science Inventory

Established microbial ecology analytical techniques for measuring the quantity and activity of bacteria were examined for use on biological granular activated carbon (GAC). ctivity was determined using the fluorescein diacetate (FDA) assay. he assay was tested and accordingly cor...

411

Ultrastructure of the zinc iodide-osmic acid stained cells in guinea pig small intestine.  

PubMed Central

This study has demonstrated that cells stained using the zinc iodide-osmic acid (ZIO) method have the same fine structural features as those of fibroblasts. They have a well developed Golgi apparatus, granular endoplasmic reticulum and many mitochondria. They have no basal lamina. They are distributed in association with the deep muscular plexus, within the outer circular muscle layer and in the space between the circular and longitudinal muscle layers. Since this staining method is believed to co-stain nerves and interstitial cells of Cajal, we concluded that these ZIO-positive, fibroblast-like cells represent at least some, if not all, of the interstitial cells which appeared in the original description by Cajal. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 PMID:7592010

Zhou, D S; Komuro, T

1995-01-01

412

Study of glasses with grisailles from historic stained glass windows of the cathedral of León (Spain)  

NASA Astrophysics Data System (ADS)

This work concerns the study of grisailles of historic glass samples from stained glass windows of the Cathedral of León, which were removed during the restoration carried out in 19th century. Both the glass samples and their coloured grisailles showed very different chemical composition and macroscopic heterogeneity. As a general rule their deterioration degree is rather moderate, maybe due to the pieces removal that preserve them from the high atmospheric pollution occurred in the last century. The present research pointed out the physical characteristics, chemical compositions and deterioration degree of the samples selected from the most important Spanish ensemble of Medieval and Renaissance stained glass windows. Moreover, this work offers sufficient results to be compared with those formerly obtained for other stained glass windows from European cathedrals and churches.

Carmona, N.; Villegas, M. A.; Navarro, J. M. Fernández

2006-06-01

413

Comparative analysis of colorimetric staining in skin using open-source software.  

PubMed

Colorimetric staining techniques such as immunohistochemistry (IHC), immunofluorescence (IF) and histochemistry (HC) provide useful information regarding the localization and relative amount of a molecule/substance in skin. We have developed a novel, straightforward method to assess colorimetric staining by combining features from two open-source software programs. As a proof of principle, we demonstrate the utility of this approach by analysing changes in skin melanin deposition during the radiation-induced tanning response of Yucatan mini-pigs. This method includes a visualization step to validate the accuracy of colour selection before quantitation to ensure accuracy. The data show that this method is robust and will provide a means to obtain accurate comparative analyses of staining in IHC/IF/HC samples. PMID:25393687

Billings, Paul C; Sanzari, Jenine K; Kennedy, Ann R; Cengel, Keith A; Seykora, John T

2015-02-01

414

The morphology of Golgi-stained neurons in lamina II of the rat spinal cord.  

PubMed Central

Golgi-stained neurons in Lamina II of the rat spinal cord were examined by light microscopy. Stalked and islet cells similar to those seen in other species were found. Stalked cells were present in large numbers in the dorsal part of the lamina where they made up nearly half the population of stained cells. Islet cells were found throughout the lamina and constituted about one third of the total population. In the ventral part of the lamina half of the stained cells did not fall into either category, but could be divided into groups on the basis of dendritic spread. The axons of many of these cells either remained in Lamina II or passed ventrally into Lamina III. Some of these cells may correspond to the stellate or the II-III border cells which have been seen in human spinal cord and cat medulla respectively. PMID:2447052

Todd, A J; Lewis, S G

1986-01-01

415

Direct isolation of genes encoded within a homogeneously staining region by chromosome microdissection.  

PubMed Central

Identification of genes involved in recurring chromosome rearrangements has provided significant insight into the molecular basis of malignancy. We describe here a strategy combining chromosome microdissection and hybrid selection for the direct isolation of chromosome region-specific genes. We modeled this strategy by using sequences recovered from the microdissection of a homogeneously staining region to allow isolation of genes that were overexpressed and present at high copy number within the homogeneously staining region, including the direct isolation of two genes encoded within a 12q homogeneously staining region found in the osteosarcoma cell line OsA-CL. Although first applied to amplified genes, this strategy should be applicable to the isolation of cDNAs from any chromosomal region. Images PMID:8090779

Su, Y A; Trent, J M; Guan, X Y; Meltzer, P S

1994-01-01

416

Enumeration of small ciliates in culture by flow cytometry and nucleic acid staining.  

PubMed

We developed a fast and simple protocol for accurate quantification of small freshwater ciliates by flow cytometry (FCM). The ciliates were stained with several nucleic acid stains such as TO-PRO-1, YO-YO-1 and PicoGreen, and analysed by a commercially available flow cytometer. The method was tested with cultures of the prostomatid species Urotricha farcta and Balanion planctonicum, including the small cryptophyte Cryptomonas sp. as food. Of the dyes tested, TO-PRO-1 gave the best results. Flow cytometric results agreed well with microscopic counts. Due to its greater speed and accuracy, FCM was superior to light microscopy. FCM was also superior to electronical particle counting and sizing (EPCS). Of particular importance, FCM in combination with TO-PRO-1 staining allowed unequivocal discrimination in cases of overlapping size distributions between the target population (i.e., the ciliate predators) and other particles (the cryptophyte prey, detritus). PMID:11830303

Lindström, Eva S; Weisse, Thomas; Stadler, Peter

2002-04-01

417

THE USE OF BISMUTH AS AN ELECTRON STAIN FOR NUCLEIC ACIDS  

E-print Network

Evidence is presented to show that bismuth combines in vitro with the phosphate of nucleic acids in a manncr similar to its reaction with inorganic phosphate. When tested under similar conditions, protein exhibited no attraction for bismuth. Thc results of the in vitro experiments, which are of interest within themselves, may be indirectly applicable to in vivo staining. Dividing cells of onion root tips were fixed in OsO4, stained with bismuth, and examined in the electron microscope. The electron opacity of cell structures known to contain nucleic acids was enhanced by bismuth, while organelles known to lack appreciable quantities of DNA or RNA showed little, if any, change. Bismuth is particularly effective as a stain for thc chromatin material during interphase and for the chromosomcs during division.

Peter Albersiieim, Ph.D.; Ursula Killias

418

Diagnostic utility of WT-1 cytoplasmic stain in variety of vascular lesions  

PubMed Central

Vascular lesions are commonly encountered in routine pathologic practice and often pose diagnostic challenges owing to their morphologic diversity. Although WT-1 expression was reported in some vascular tumors, little is known about its staining patterns in a spectrum of vascular lesions from various locations. We examined WT-1 immunostain in 95 cases of vascular lesions including angiosarcomas (AS, 19 cases), hemangioendotheliomas (HE, 5), Kaposi’s sarcomas (KS, 4), cavernous hemangiomas (CVH, 12), capillary hemangiomas (CPH, 7), pyogenic granulomas (PG, 4), lymphangiomas (LA, 4), hemangiopericytomas (HP, 5), glomus tumors (GT, 8), vascular malformation (VM, 13) and granulation tissue (GRT, 14). Strong WT-1 cytoplasmic stain was invariably observed in all cases of malignant and borderline vascular tumors including AS (19/19), KS (4/4) and HE (5/5). WT-1 was also consistently expressed in CPH (7/7), PG (4/4), and GRT (14/14), while it became weaker in VM (10/13) and often negative in CVH (2/12) and LA (0/4). WT1 stain was not demonstrated in HP (0/5) and rarely in GT (2/8). We conclude that consistent and diffuse WT-1 cytoplasmic stain in AS, HE and KS can be useful in distinguishing these tumors from poorly differentiated tumors with mimicking features. On the other hand, reliable WT-1 stain in CPH, PG and GRT may help in differential diagnosis with non-endothelial vascular tumors such as GT and HP. Recognizing the WT-1 cytoplasmic stain in a broad spectrum of benign and neoplastic tissues is critical in formulating appropriate immunohistochemical panels and avoiding misinterpretation of results. PMID:24966966

Galfione, Sarah K; Ro, Jae Y; Ayala, Alberto G; Ge, Yimin

2014-01-01

419

Rapid on-site cytologic examination of 1500 breast lesions using the modified Shorr's stain.  

PubMed

BACKGROUND: Rapid on-site evaluation (ROSE) cytology enables sample quality assessment in the procedure room and facilitates the process of examination. While its use for mammary lesions in one-stop breast clinics has been reported, its usefulness as a cytologic diagnostic tool has not been fully explored. METHODS: A total of 1500 examinations of core-needle biopsy imprint/fine-needle aspiration cytology were performed for outpatients with breast lesions. The slides were immediately processed with modified Shorr's stain, which can be completed within a few minutes yet produces specimens of similar staining quality as the Papanicolaou (Pap) stain. The adequacy of sampling was evaluated on site, and a cytologic diagnosis was also made. ROSE cytologic findings were classified into five grades: class 1, inadequate; class 2, benign; class 3, indeterminate; class 4, suspicious for malignancy; class 5, malignant. If enough epithelial cells could not be obtained despite repeated examinations, the sample was scored as ineligible. These scores were utilized for patient management. Final cytologic diagnoses were made with conventional Pap stains. RESULTS: Reproducibility of scores between both staining methods was excellent (weighted ? statistic = 0.985). When compared class by class, concordance of cytologic diagnoses was particularly high in class 2 and 5 Shorr scores, in which the agreement with Pap diagnoses was 92.8 and 93.6 %, respectively. CONCLUSIONS: Our modified Shorr's staining protocol was useful to reduce the time for the diagnosis and treatment planning of breast lesions suspected of being breast cancer. It is beneficial for both the patients and clinicians. PMID:23733595

Sakuma, Takahiko; Mimura, Akihiro; Tanigawa, Naoto; Takamizu, Ryuichi; Morishima, Hirotaka; Matsunami, Nobuki

2013-06-01

420

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

PubMed Central

Methods are presented for the staining of blocks of tissue with trivalent indium so that good contrast and good specificity for nucleic acids is achieved for the electron microscope. The tissue is fixed in organic fixative, dehydrated, subjected to reduction by lithium borohydride, acetylated by acetic anhydride, stained with trivalent indium dissolved in organic solvent, and embedded. The embedding material may be either Vestopal or butyl methacrylate especially handled to eliminate the "explosion" phenomenon. Numerous new problems encountered are discussed and a brief description of the findings is included. PMID:14005301

Watson, Michael L.; Aldridge, William G.

1961-01-01

421

Electron staining of the cell surface coat by osmium-low ferrocyanide  

Microsoft Academic Search

In aldehyde-fixed liver and renal cortex of rat and mouse several variations of postfixation with osmium tetroxide plus potassium ferrocyanide (FeII) were tried. Depending on the ferrocyanide concentration different staining patterns were observed in TEM.-Osmium-High Ferrocyanide [40 mM (~1%) OsO4+36 mM (~1.5%) FeII, pH 10.4], stains membranes and glycogen. Cytoplasmic ground substance, mitochondrial matrices and chromatin are partially extracted, cell

W. F. Neiss

1984-01-01

422

Fast and automatic imaging of immunoenzyme-stained neuronal circuits in the whole brain of Drosophila  

NASA Astrophysics Data System (ADS)

Knowledge of neuronal wiring and morphogenesis in Drosophila is essential to understand brain function and dysfunction. The immunoenzyme method based on horseradish peroxidase/diaminobenzidine (HRP/DAB) provides high-contrast images to resolve details underlying neuronal architecture. However, the poor staining penetration and a lack of corresponding three-dimensional imaging methodology limit its application. Herein, we modified the HRP/DAB method to stain neuronal circuits in the whole brain of Drosophila. Furthermore, we found that imaging with the micro-optical sectioning tomography system provided a fast and automatic method that could dissect cell-specific neuroanatomical architecture at a submicron voxel resolution.

Tian, Qingping; Yuan, Jing; Li, Yuxin; Jiang, Tao; Gong, Hui; Zhou, Wei

2014-09-01

423

Single step modified ink staining for Tzanck test: quick detection of herpetic giant cells in Tzanck smear.  

PubMed

Tzanck test has been recently re-evaluated as a method for the diagnosis of herpes virus infection. Giemsa staining for the Tzanck test is time-consuming and laborious. There is a need to develop simple and quick staining methods for bedside diagnosis of this disease. We report a single step and quick method for staining herpes giant cells in Tzanck smears using routinely available inks and physiological saline. A keratinocyte cell line (HaCaT) was cultured on a slide glass and stained with various commercially available blue, blue-black and black inks serially diluted with physiological saline. Clinical smear samples from herpes lesions were also stained with these solutions without specific pretreatment. The nuclei of HaCaT were clearly stained showing high contrast with the cytoplasm using 5% Parker-Quink blue-black ink saline solution. Concentration of ink solution higher or lower than 5% resulted in less contrast. Blue or black inks or other manufacturers' inks can also be used, but staining of the cultured keratinocytes was less clear. Smear of clinical samples from herpes lesions were also stained with 5% ink solution. The nuclei of the multinucleated giant cells were clearly stained, and the sample could be immediately used for microscopic examination. One step staining of Tzanck smear using this diluted ink solution is an inexpensive and a convenient bedside diagnostic tool for the dermatologist. PMID:21951152

Mizutani, Hitoshi; Akeda, Tomoko; Yamanaka, Kei-Ichi; Isoda, Kenichi; Gabazza, Esteban C

2012-02-01

424

Gram Stain  

MedlinePLUS

... identify the general type of bacteria or sometimes fungi ( microorganisms ) in a sample taken from the site ... of an infection; a sample of bacteria or fungi grown and isolated in culture Test Preparation Needed? ...

425

Fluorescein Dye Penetration in Round Top Rhyolite (Hudspeth County, Texas, USA) to Reveal Micro-permeability and Optimize Grain Size for Heavy REE Heap Leach  

NASA Astrophysics Data System (ADS)

Millimeter- and micrometer-scale permeability of fine-grained igneous rocks has generated limited research interest. Nonetheless, the scale and distribution of such micro-permeability determines fluid penetration and pathways, parameters that define both the ability to heap leach a rock and the optimal grain size for such an operation. Texas Rare Earth Resources is evaluating the possibility of heap leaching of yttrium and heavy rare earth elements (YHREE) from the peraluminous rhyolite laccolith that forms one-mile-diameter Round Top Mountain. The YHREEs in this immense, surface-exposed deposit (minimum 1.6 billion tons, Texas Bureau Economic Geology) are dilute and diffuse, suggesting leaching as the best option for recovery. The REE grade is 0.05% and YHREEs comprise more than 70% of the total REE content. The YHREEs are hosted exclusively in micron-scale yttrofluorite grains, which proved soluble in dilute sulfuric acid. Laboratory experiments showed YHREE recoveries of up to 90%. Within limits, recoveries decrease with larger grain sizes, and increase with acid strength and exposure time. Our research question centers on dissolution effectiveness: Is YHREE recovery, relative to grain size, limited by (1) diffusion time of acid into, and dissolved solids, including YHREEs, out of the micro-permeability paths inherent in the rock particles; (2) the effective lengths of the natural micro-permeability paths in the rock; or (3) the putative role of the acid in dissolving new micro-paths into the grains? The maximum grain size should not exceed twice the typical path length (unless acid creates new paths), lest YHREEs in the core of a larger grain than that not be reached by acid. If instead diffusion time is limiting, longer leach time may prove effective. Rather than perform an extensive and expensive series of laboratory leaching experiments--some of which would be several months in duration--to determine optimal grain size, we developed a technique to efficiently determine the limits of penetration of water into the rhyolite. We cut parallel-sided slabs of Round Top rhyolite at staged thickness up to 10 mm. We then wet one side and view the opposite side over time under UV light to detect breakthrough of the fluorescein dye. Because of its extremely low visual detection limits, well below the ppm level, the dye has been widely used in biochemical research, as a tracer in surface and ground water studies, in delineating invisible cracks in such structural material as motor blocks, and in detecting corneal abrasions. We have been successful in detecting breakthrough at different rhyolite thicknesses. Continuing studies focus on mapping of the 2-dimensional distribution of the permeability via hand lens and low-power microscope; use of visible light dyes; and examination of specimens pre- and post-acid leaching to determine whether acid exposure produced significant new micro-permeability.

Negron, L. M.; Clague, J. W.; Gorski, D.; Amaya, M. A.; Pingitore, N. E.

2013-12-01

426

2-(2-Hydroxy-5-nitrobenzylidene)-1,3-indanedione versus Fluorescein Isothiocyanate in Interaction with Anti-hFABP Immunoglobulin G1: Fluorescence Quenching, Secondary Structure Alteration and Binding Sites Localization.  

PubMed

The first step in determining whether a fluorescent dye can be used for antibody labeling consists in collecting data on its physical interaction with the latter. In the present study, the interaction between the 2-(2-hydroxy-5-nitrobenzylidene)-1,3-indanedione (HNBID) dye and the IgG1 monoclonal mouse antibody anti-human heart fatty acid binding protein (anti-hFABP) has been investigated by fluorescence and circular dichroism spectroscopies and complementary structural results were obtained by molecular modeling. We have determined the parameters characterizing this interaction, namely the quenching and binding constants, classes of binding sites, and excited state lifetimes, and we have predicted the localization of HNBID within the Fc region of anti-hFABP. The key glycosidic and amino acid residues in anti-hFABP interacting with HNBID have also been identified. A similar systematic study was undertaken for the well-known fluorescein isothiocyanate fluorophore, for comparison purposes. Our results recommend HNBID as a valuable alternative to fluorescein isothiocyanate for use as a fluorescent probe for IgG1 antibodies. PMID:23434655

Stan, Dana; Mihailescu, Carmen-Marinela; Savin, Mihaela; Matei, Iulia

2013-01-01

427

Combined laser and surgical treatment of giant port wine stain malformation - Case report  

NASA Astrophysics Data System (ADS)

Background:Port-wine stains (PWS) are vascular malformations of the skin concerning about 0,3% of the population. Though various laser systems have been used for various treatment regimens the treatment of PWS of large size is especially difficult and demanding from aesthetic and psychological point of view.

Siewiera, I.; Drozdowski, P.; Wójcicki, P.

2012-10-01

428

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

E-print Network

colloidal Coomassie blue, while ruthenium complexes have a sensitivity slightly inferior to that of silver1 ABOUT THE MECHANISM OF INTERFERENCE OF SILVER STAINING WITH PEPTIDE MASS SPECTROMETRY Sophie, DRDC/BECP CEA-Grenoble, 17 rue des martyrs, F-38054 GRENOBLE CEDEX 9, France (Running title): silver

Paris-Sud XI, Université de

429

Quantitative analysis of stain variability in histology slides and an algorithm for standardization  

NASA Astrophysics Data System (ADS)

This paper presents data on the sources of variation of the widely used hematoxylin and eosin (H&E) histological staining, as well as a new algorithm to reduce these variations in digitally scanned tissue sections. Experimental results demonstrate that staining protocols in different laboratories and staining on different days of the week are the major factors causing color variations in histopathological images. The proposed algorithm for standardizing histology slides is based on an initial clustering of the image into two tissue components having different absorption characteristics for different dyes. The color distribution for each tissue component is standardized by aligning the 2D histogram of color distribution in the hue-saturation-density (HSD) model. Qualitative evaluation of the proposed standardization algorithm shows that color constancy of the standardized images is improved. Quantitative evaluation demonstrates that the algorithm outperforms competing methods. In conclusion, the paper demonstrates that staining variations, which may potentially hamper usefulness of computer assisted analysis of histopathological images, can be reduced considerably by applying the proposed algorithm.

Ehteshami Bejnordi, Babak; Timofeeva, Nadya; Otte-Höller, Irene; Karssemeijer, Nico; van der Laak, Jeroen A. W. M.

2014-03-01

430

Evaluation of maturity group III soybean lines for resistance to purple seed stain in Mississippi, 2010  

Technology Transfer Automated Retrieval System (TEKTRAN)

Purple seed stain (PSS) of soybean is an important disease caused by Cercospora kikuchii. PSS reduces seed quality and market grade, affects seed germination and vigor, and has been reported wherever soybeans are grown worldwide. In 2009, PSS caused 6.4 million bushels of yield losses in 16 southern...

431

Improved selective, simple, and contrast staining of acidophilic neurons with vanadium acid fuchsin  

Microsoft Academic Search

Acidophilia is one of the hallmarks of acute neuronal damage and death in brain ischemia, excitotoxic and traumatic lesions and epileptic seizures. We here describe a novel and simple method for visualizing acidophilic neurons on paraffin sections, using vanadium acid fuchsin (VAF) staining and toluidine blue or hematoxylin counterstaining. Paraffin sections of the brain fixed in ethanol–formalin–acetic acid mixture are

Ilya V Victorov; Konstantin Prass; Ulrich Dirnagl

2000-01-01

432

Development of an Automated Microscope for Supporting Qualitative Asbestos Analysis by Dispersion Staining  

Microsoft Academic Search

This paper introduces automated microscopic obser- vation supporting qualitative asbestos analysis. Visual qualitative asbestos evaluation generally involves dis- persion staining. Operators conventionally check and count asbestos fibers visually by microscope. We are developing automated microscopic observation to sup- port qualitative asbestos analysis. The system images fibers by microscope and saves them automatically to a database. We introduce system concepts and

Kuniaki Kawabata; Soichiro Morishita; Kazuhiro Hott; Taketoshi Mishima; Hiroshi Mizoguchi; Haruhisa Takahashi

2009-01-01

433

Development of a polarized microscopic image management system for supporting asbestos qualitative analysis utilizing dispersion staining  

Microsoft Academic Search

This paper describes development of an automated polarized microscopic image management system for supporting qualitative analysis of asbestos. Dispersion staining is a visual observation method. Experts count all particles in the microscope view and also the number of the fibrous asbestos fibers. For supporting this work, we are developing an automated system to ease experts' burdens for efficient observation. The

Y. Tsubota; K. Kawabata; H. Yamazaki; K. Hotta; H. Asama; H. Mizoguchi; H. Takahashi; T. Mishima

2009-01-01

434

AN EVALUATION OF THREE CONVENTIONAL HISTOLOGICAL TECHNIQUES FOR STAINING THE CERATA OF CRATENA PILATA  

EPA Science Inventory

Fixation and staining methods for different types of tissue in the marine nudibranch Cratena pilata were evaluated. Cratena pilata, a marine snail in the Phylum Mollusca, has the ability to take stinging cells, called nematocysts, from ingested animals belonging to another phylum...

435

CREST staining of micronuclei from free-living rodents to detect environmental contamination in situ  

Microsoft Academic Search

In this work immunofluorescent antikinetochore (CREST) staining was used to analyse bone marrow micronuclei (MN) from free-living animals belonging to four different rodent species. Yellow-necked mice (Apodemus flavicollis ) and bank voles (Clethrionomys glareolus) were trapped in the Czech Republic, Algerian mice (Mus spretus) in Spain and house mice (Mus musculus domesticus) in Italy. Animals were collected in areas displaying

Francesca Degrassi; Caterina Tanzarella; Luisa Anna Ieradi; Jan Zima; Anna Cappai; Antonella Lascialfari; Fabio Allegra; Mauro Cristaldi

1999-01-01

436

Validation of quantitative susceptibility mapping with Perls' iron staining for subcortical gray matter.  

PubMed

Quantitative susceptibility mapping (QSM) measures bulk susceptibilities in the brain, which can arise from many sources. In iron-rich subcortical gray matter (GM), non-heme iron is a dominant susceptibility source. We evaluated the use of QSM for iron mapping in subcortical GM by direct comparison to tissue iron staining. We performed in situ or in vivo QSM at 4.7 T combined with Perls' ferric iron staining on the corresponding extracted subcortical GM regions. This histochemical process enabled examination of ferric iron in complete slices that could be related to susceptibility measurements. Correlation analyses were performed on an individual-by-individual basis and high linear correlations between susceptibility and Perls' iron stain were found for the three multiple sclerosis (MS) subjects studied (R(2) = 0.75, 0.62, 0.86). In addition, high linear correlations between susceptibility and transverse relaxation rate (R2*) were found (R(2) = 0.88, 0.88, 0.87) which matched in vivo healthy subjects (R(2) = 0.87). This work validates the accuracy of QSM for brain iron mapping and also confirms ferric iron as the dominant susceptibility source in subcortical GM, by demonstrating high linear correlation of QSM to Perls' ferric iron staining. PMID:25462797

Sun, Hongfu; Walsh, Andrew J; Lebel, R Marc; Blevins, Gregg; Catz, Ingrid; Lu, Jian-Qiang; Johnson, Edward S; Emery, Derek J; Warren, Kenneth G; Wilman, Alan H

2015-01-15

437

Differential staining of acid glycosaminoglycans (mucopolysaccharides) by Alcian blue in salt solutions  

Microsoft Academic Search

The application of the “critical electrolyte concentration” (CEC) concept to the differentiation of acidic glycosaminglycans (mucopolysaccharides) is described. Alcian Blue 8GX stains with increasing selectivity as increasing amounts of magnesium chloride are incorporated into the dye solution. Model experiments with pure polyanions, or artifically carboxylated, phosphorylated and sulphated liver sections, showed that binding of dye to carboxylate or phosphate groups

J. E. Scott; J. Dorling

1965-01-01

438

The Polarization of Fluorescence of DNA Stains Depends on the Incorporation Density of the  

E-print Network

The Polarization of Fluorescence of DNA Stains Depends on the Incorporation Density of the Dye: The fluorescence induced by polarized light sources, such as the lasers that are used in flow cytometry, is often polarized and anisotropic. In addition, most optical detector systems are sensitive to the direc- tion

Asbury, Chip

439

ASSESSING NEZARA VIRIDULA (HEMIPTERA: PENTATOMIDAE) FEEDING DAMAGE IN MACADAMIA NUTS BY USING A BIOLOGICAL STAIN  

Technology Transfer Automated Retrieval System (TEKTRAN)

Damage caused by Nezara viridula to macadamia nuts is normally determined after nuts are harvested and processed, which may be many months after damage occurred in the field. We developed a method using ruthenium red dye to stain stink bug feeding probes and indirectly assess feeding activity in mac...

440

Blockface Histology with Optical Coherence Tomography: A Comparison with Nissl Staining  

PubMed Central

Spectral domain optical coherence tomography (SD-OCT) is a high resolution imaging technique that generates excellent contrast based on intrinsic optical properties of the tissue, such as neurons and fibers. The SD-OCT data acquisition is performed directly on the tissue block, diminishing the need for cutting, mounting and staining. We utilized SD-OCT to visualize the laminar structure of the isocortex and compared cortical cytoarchitecture with the gold standard Nissl staining, both qualitatively and quantitatively. In histological processing, distortions routinely affect registration to the blockface image and prevent accurate 3D reconstruction of regions of tissue. We compared blockface registration to SD-OCT and Nissl, respectively, and found that SD-OCT-blockface registration was significantly more accurate than Nissl-blockface registration. Two independent observers manually labeled cortical laminae (e.g. III, IV and V) in SD-OCT images and Nissl stained sections. Our results show that OCT images exhibit sufficient contrast in the cortex to reliably differentiate the cortical layers. Furthermore, the modalities were compared with regard to cortical laminar organization and showed good agreement. Taken together, these SD-OCT results suggest that SD-OCT contains information comparable to standard histological stains such as Nissl in terms of distinguishing cortical layers and architectonic areas. Given these data, we propose that SD-OCT can be used to reliably generate 3D reconstructions of multiple cubic centimeters of cortex that can be used to accurately and semi-automatically perform standard histological analyses. PMID:24041872

Magnain, Caroline; Augustinack, Jean C.; Reuter, Martin; Wachinger, Christian; Frosch, Matthew P.; Ragan, Timothy; Akkin, Taner; Wedeen, Van J.; Boas, David A.; Fischl, Bruce

2015-01-01

441

Suppurative keratitis in rural Bangladesh: the value of Gram stain in planning management  

Microsoft Academic Search

External eye diseases which result in corneal scarring are an important cause of blindness in Bangladesh and at the Chittagong Eye Infirmary and Training Complex (EITC) over 200 cases of suppurative keratitis are managed each year. We reviewed the records of 127 cases of microbial keratitis to determine the relative contributions of Gram stain and culture to diagnosis of the

G. Williams; K. McClellan; F. Billson

1991-01-01

442

Reaction of maturity group V soybean lines to purple seed stains in Mississippi 2010  

Technology Transfer Automated Retrieval System (TEKTRAN)

In 2009, soybean purple seed stain (PSS) caused 6.4 million bushels of yield losses in 16 southern states. This disease severely reduces seed market grade and affects seed germination and vigor. PSS is caused by Cercospora kikuchii and is an economy important disease. To identify new sources of resi...

443

Evaluation of maturity group IV soybean lines for resistance to purple seed stains in Mississippi 2010  

Technology Transfer Automated Retrieval System (TEKTRAN)

Purple seed stain (PSS) of soybean is an important disease caused by Cercospora kikuchii. PSS reduces seed quality and market grade, affects seed germination and vigor, and has been reported wherever soybeans are grown worldwide. In 2009, PSS caused 6.4 million bushels of yield losses in 16 southern...

444

Sensitivity and specificity of epithelial membrane antigen staining patterns in ependymomas  

Microsoft Academic Search

Pattern and extent of epithelial membrane antigen (EMA) immunoreactivity in ependymomas as compared to other glial tumors have only been investigated in small series. To determine sensitivity and specificity of EMA staining, 54 ependymomas were evaluated in comparison to 54 glioblastomas, 43 fibrillary astrocytomas and 21 oligodendrogliomas. Distinct punctate intracytoplasmic EMA immunoreactivity was observed in 48\\/54 ependymomas (89%), whereas ring-like

Martin Hasselblatt; Werner Paulus

2003-01-01

445

Design Implications from a Usability Study of GramStain-Tutor.  

ERIC Educational Resources Information Center

Describes a usability study conducted with health sciences students at the University of Washington that explored interface issues in the GramStain Tutor, an educational software program on CD-ROM, particularly the navigation of the program and the use of embedded design features. (LRW)

Kim, Sara; Brock, Douglas; Orkand, Adam; Astion, Michael

2001-01-01

446

Iodamoeba butschlii in an anal pap test confirmed by iodine stain.  

PubMed

We report the finding of Iodamoeba butschlii amebic cysts on a liquid-based anal Pap smear from an HIV-positive male. Iodine staining of the smear confirmed the diagnosis. It is important to distinguish I. butschlii from pathogenic ameobae and other organisms seen on anal Pap smears. PMID:24167099

Schuetz, Audrey N; Pritt, Bobbi S; Schreiner, Andrew M

2014-09-01

447

Microwave-enhanced ink staining for fast and sensitive protein quantification in proteomic studies.  

PubMed

A novel microwave-enhanced ink staining method was developed for rapid and sensitive estimation of protein content in sample buffers containing chaotropes, dyes, detergents, and reducing agents. Dye-based Blue-Black ink was used to quantitatively visualize proteins spotted on a nitrocellulose membrane. The total staining time was greatly reduced to 3 min by brief exposure to microwave radiation. The stained membrane was washed with distilled water, baked in a microwave oven for complete desiccation, transparentized with mineral oil, and documented by a desktop scanner or densitometer. Only 1 microL of protein sample (protein solubilized in SDS-PAGE sample buffer or IEF rehydration buffer) was used for protein spotting. The novel solid-phase protein assay gives a 500-fold dynamic range from 19.5 to 10000 ng/microL and can be scaled up for high-throughput protein quantification analysis. The fast, sensitive and low-cost microwave-enhanced ink staining procedure is ideal for protein quantification in proteomic analysis. PMID:17203983

Wu, Xue-Ping; Cheng, Yong-Sheng; Liu, Jin-Yuan

2007-01-01

448

Intraoperative immunohistochemistry staining of sentinel nodes in breast cancer: clinical and economical implications.  

PubMed

The study aimed to evaluate intraoperative immunohistochemistry (IHC) staining of sentinel nodes in primary breast cancer surgery. We analysed retrospectively 1209 consecutive sentinel node procedures and compared the rate of late positive metastases in sentinel node biopsy (SNB) and the duration of the surgical procedures before (n=706) and after (n=503) introducing intraoperative IHC on frozen section. We also did a cost analysis. Intraoperative IHC staining led to a lowering of the late positive SNB rate. Introducing IHC gave a decrease in the late positive rate from 93 to 52% (p<0.0001) for isolated tumour cell metastasis, from 56 to 36.4% (p<0.02) for micrometastasis, and from 16 to 5% (p<0.01) for macrometastasis. The surgical procedures were slightly prolonged for lumpectomies but not for mastectomies after introducing intraoperative IHC staining. The cost analysis showed an overall cost saving of approximately 40%. In conclusion, intraoperative IHC staining of the SNB lowered the late positive rate and gave an overall cost saving. PMID:18490162

Holm, Marianne; Paaschburg, Birgitte; Balslev, Eva; Axelsson, Christen Kirk; Willemoe, Gro Linno; Flyger, Henrik Lavlund

2008-08-01

449

Meconium-stained amniotic fluid and its association with obstetric infections  

Microsoft Academic Search

Meconium-stained amniotic fluid (MSAF) complicates the intrapartum course of 1.5% to 18% of pregnancies. In addition to predisposing to the meconium aspiration syndrome, MSAF is also an established risk factor for neonatal sepsis and for intrapartum and postpartum maternal infection. Meconium enhances the growth of bacteria in amniotic fluid by serving as a growth factor, inhibits the bacteriostatic properties of

Rodney K Edwards

1998-01-01

450

Detecting dentinal sclerosis in decalcified sections with the Pollak trichrome connective tissue stain.  

PubMed

The supply of young human teeth for controlled human pulp studies is inadequate. Dentinal sclerosis in older teeth modifies the expected pulpal responses. Decalcification, necessary to interpret pulpal responses, destroys the primary evidence of sclerosis. At present we are unable to detect the degree of sclerosis and the pulp response in the same preparation. Valid interpretations on older teeth cannot be accomplished until this problem is resolved. Fifty-seven teeth, normal and carious, representing a wide age span were fixed, embedded in Bioplastic and sectioned with the Bronwill Model No. 77 through the middle of the M-D plane on the labio-lingual plane to obtain an undecalcified ground section for examination with reflected and transmitted light and Faxitron 805 microradiography. The remaining portions of the tooth were decalcified, processed in paraffin and subjected to a variety of histochemical reactions including the Pollak trichrome stain. The Pollak trichrome and the Pollak trichrome solution No. 6 modification stains provided the greatest correlations with the sclerotic areas in the radiographs of the ground sections. Areas of dentin indicative of sclerosis prior to decalcification stained orange-red with the Pollak trichrome stain and deep orange with the Pollak trichrome solution No. 6 modification. PMID:6164773

Stanley, H R; Broom, C A; Spiegel, E H; Schultz, M S

1980-11-01

451

CHARACTERIZATION OF ORGANIC EMISSIONS FROM A WOOD FINISHING PRODUCT - WOOD STAIN  

EPA Science Inventory

The paper gives results of the measurement of emission characteristics of four organic compounds (nonane, decane, undecane, and 1,2,4-trimethylbenzene) from a wood finishing product, wood stain, in an environmental chamber. It was found that the emission patterns of the four orga...

452

A hematoxylin staining technique to locate sites of aluminum binding in aquatic plants and animals  

Microsoft Academic Search

This paper describes a modified hematoxylin staining technique that can be used to locate sites of A1 binding in freshwater plants and animals. This technique is fast, simple, and inexpensive to use. It is more reliable for organisms raised under controlled conditions, although it can be used on organisms isolated from the field. In the presence of Al, a purple

Magda Havas

1986-01-01

453

Survey and Detection of Endophytic Fungi in Lolium Germ Plasm by Direct Staining and Aphid Assays  

Microsoft Academic Search

Wilson, A. D., Clement, S. L., and Kaiser, W. J. 1991. Survey and detection of endophytic fungi in Lolium germ plasm by direct staining and aphid assays. Plant Dis. 75:169-173. Clavicipitaceous anamorphic endophytes were detected in 28 of 85 accessions from five of eight species in a collection of Lolium germ plasm. Comparative descriptions of endophytic mycelium in seeds of

A. DAN WILSON; STEPHEN L. CLEMENT; WALTER J. KAISER

454

Efficacy of baking soda-containing chewing gum in removing natural tooth stain.  

PubMed

A 14-week, double-blind, randomized clinical trial was conducted with 126 healthy volunteers to compare the efficacy of twice-daily use of 3 baking soda-containing chewing gums in removing natural tooth stain when used in conjunction with a program of regular oral hygiene. All 3 chewing gums significantly reduced extrinsic stain (P < .0001) and improved the whitened appearance of teeth (P < .0001) at both the 2-week interim and the final 4-week evaluations. ARM & HAMMER DENTAL CARE The Baking Soda Gum (AHDC) reduced dental stain by 70.8%, compared to reductions of 71.9% and 65.3%, after use of 2 experimental gum formulations. Whitened appearance improved by 1.73 shade tabs using AHDC gum, and up to 2.49 shade tabs with the experimental formulations. These results suggest that the use of baking soda-containing gum after meals, in conjunction with good oral hygiene, can improve both extrinsic dental staining and the whitened appearance of teeth. PMID:11913307

Mankodi, S M; Conforti, N; Berkowitz, H

2001-07-01

455

Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections  

Microsoft Academic Search

Synopsis  Sirius Red, a strong anionic dye, stains collagen by reacting, via its sulphonic acid groups, with basic groups present in the collagen molecule. The elongated dye molecules are attached to the collagen fibre in such a way that their long axes are parallel. This parallel relationship between dye and collagen results in an enhanced birefringency.Examination of tissue sections from 15

L. C. U. Junqueira; G. Bignolas; R. R. Brentani

1979-01-01

456

Enhanced visualization of microbial biofilms by staining and environmental scanning electron microscopy.  

PubMed

Bacterial biofilms, i.e. surface-associated cells covered in hydrated extracellular polymeric substances (EPS), are often studied with high-resolution electron microscopy (EM). However, conventional desiccation and high vacuum EM protocols collapse EPS matrices which, in turn, deform biofilm appearances. Alternatively, wet-mode environmental scanning electron microscopy (ESEM) is performed under a moderate vacuum and without biofilm drying. If completely untreated, however, EPS is not electron dense and thus is not resolved well in ESEM. Therefore, this study was towards adapting several conventional SEM staining protocols for improved resolution of biofilms and EPS using ESEM. Three different biofilm types were used: 1) Pseudomonas aeruginosa unsaturated biofilms cultured on membranes, 2) P. aeruginosa cultured in moist sand, and 3) mixed community biofilms cultured on substrates in an estuary. Working with the first specimen type, a staining protocol using ruthenium red, glutaraldehyde, osmium tetroxide and lysine was optimized for best topographic resolution. A quantitative image analysis tool that maps relief, newly adopted here for studying biofilms, was used to compare micrographs. When the optimized staining and ESEM protocols were applied to moist sand cultures and aquatic biofilms, the smoothening effect that bacterial biofilms have on rough sand, and the roughening that aquatic biofilms impart on initially smooth coupons, were each quantifiable. This study thus provides transferable staining and ESEM imaging protocols suitable for a wide range of biofilms, plus a novel tool for quantifying biofilm image data. PMID:17196692

Priester, John H; Horst, Allison M; Van de Werfhorst, Laurie C; Saleta, José L; Mertes, Leal A K; Holden, Patricia A

2007-03-01

457

Staining of oral epithelium with the zinc iodide-osmium reaction.  

PubMed

Some of the parameters affecting the staining of keratinized oral epithelium with the zinc iodide-osmium reaction were examined using light and electron microscopy and electron probe microanalysis. Factors examined were block size, incubation temperature and the effect of aldehyde prefixation. Large blocks (4 mm cube) were subdivided after incubation and the staining of the centre and edge compared. Generally the reaction was more variable at the edge than in the centre. Small block (1 mm cube) showed a more intense reaction when incubated at 24 degrees C than at 4 degrees C. In all these preparations, final reaction product was seen over Golgi systems, lysosome-like bodies, membrane-coating granules and, in the more intensely stained regions, over endoplasmic reticulum and nuclear membranes as well. In prefixed material, mitochondria were frequently stained in addition to the other organelles. Energy dispersive analysis showed the reaction product to be similar in all preparations and to contain high levels of zinc and osmium but not iodine. PMID:6164672

Ashrafi, S H; Squier, C A; Meyer, J

1981-01-01

458

The utility of Brilliant Cresyl Blue (BCB) staining of mammalian oocytes used for in vitro embryo production (IVP).  

PubMed

The present article summarizes the results of experiments investigating the Brilliant Cresyl Blue (BCB) staining for selection of immature oocytes before in vitro embryo production or somatic cell nuclear transfer. Developmental competence of oocytes stained with BCB and quality of blastocysts derived from such oocytes as well as the expression of apoptosis-related genes, mitochondrial DNA (mtDNA) replication-related genes and the transcripts encoded by the mitochondrial genome in BCB stained oocytes are discussed. PMID:24011188

Opiela, Jolanta; K?tska-Ksi??kiewicz, Lucyna

2013-09-01

459

Dual fluorophore doped silica nanoparticles for cellular localization studies in multiple stained cells.  

PubMed

Fluorescently labeled nanoparticles (NPs) are used in a wide range of biomedical and nanotoxicological studies to elucidate their interactions with cellular components and their intracellular localization. As commonly used fluorescence microscopes are usually limited in their performance to a few channels which detect the emitted fluorescence light in the red, green and blue color range, the simultaneous colocalization of accumulated fluorescent NPs with cellular markers is often difficult and remains a challenge due to spectral overlay of NP fluorescence and fluorescence of stained cellular components. To overcome this problem we have synthesized three different photostable dual-labeled fluorescent core/shell silica NPs with high fluorescence intensity and well-defined shape, size and surface chemistry. The synthesis route of dual fluorophore doped silica (DFDS) NPs was based on a water-in-oil microemulsion method and includes the separate incorporation of two fluorophores in the core or shell. The suitability of DFDS for colocalization studies was assessed and successfully demonstrated with human osteoblast cells. Parallel visualization of DFDS NPs with two separate microscope channels allowed cellular NP uptake and discrimination from fluorescently stained cellular components, even in triple stained cells that show fluorescence for the cytoskeleton protein actin (green), the nucleus (blue) and collagen (red). Our results demonstrate the feasibility and straightforwardness of the approach for colocalization studies at a single-cell level to discern clearly the accumulation of NPs from triple-stained cellular components. Such NPs with multiple fluorescence characteristics have a great potential to replace single fluorescent NPs for in vitro studies, when multiple staining of cellular components is required. PMID:25463504

Shahabi, Shakiba; Treccani, Laura; Dringen, Ralf; Rezwan, Kurosch

2015-03-01

460

How reliable is immunohistochemical staining for DNA mismatch repair proteins performed after neoadjuvant chemoradiation?  

PubMed

Immunohistochemistry (IHC) testing for mismatch repair proteins (MMRP) is currently being used primarily in colorectal cancer resection specimens. We aimed to compare the results of IHC staining performed on biopsy specimens obtained at endoscopy with that performed on surgical specimens after neoadjuvant therapy. Thirty-two rectal cancer subjects had paired preneoadjuvant and postneoadjuvant tissue available for IHC staining (MLH1, MSH2, MSH6, and PMS2), whereas 39 rectosigmoid cancer patients who did not receive neoadjuvant treatment served as controls. Each slide received a qualitative (absent, focal, and strong) and quantitative score (immunoreactivity [0-3] × percent positivity [0-4]). The quantitative scores of MMRP from the operative material were significantly lower in the neoadjuvant group than in the control (P < .05 for all).The scores of all MMRP from endoscopic biopsies were not significantly different between the neoadjuvant and the control groups. Disagreement between the endoscopic biopsy and the operative material was evident in 23 of 128 stains (18.5%) in the neoadjuvant group and in 12 of 156 stains (7.7%) in the control group (P = .009). In the neoadjuvant group, a disagreement pattern of "endoscopic strong operative focal" was observed in 28.1% for PMS2, 12.5% for MSH6, 12.5% for MLH1, and 6.3% for MSH2, and in the control group, this same disagreement pattern was found in 12.8% for PMS2, 7.7% for MSH6, 7.7% for MLH1, and 0% for MSH2. Based on our findings, we suggest that for rectal cancer, the endoscopic material rather than the operative material should serve as the primary material for IHC staining. PMID:25150747

Vilkin, Alex; Halpern, Marisa; Morgenstern, Sara; Brazovski, Eli; Gingold-Belfer, Rachel; Boltin, Doron; Purim, Ofer; Kundel, Yulia; Welinsky, Sara; Brenner, Baruch; Niv, Yaron; Levi, Zohar