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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...


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

PubMed Central

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

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



Dark survival strategies in marine phytoplankton assessed by cytometric measurement of metabolic activity with fluorescein diacetate  

Microsoft Academic Search

Cytometric quantification of cellular fluorescence upon cleavage of fluorescein diacetate (FDA) is presented as a sensitive\\u000a and rapid technique to assess phytoplankton metabolic activity during exposure to prolonged darkness of 10 to 12?d. Two distinct\\u000a types of metabolic response to darkness are distinguished: Type I cells (Brachiomonas submarina, Pavlova lutheri, Chrysochromulina hirta) adapt to prolonged darkness by reducing their metabolism

F. J. Jochem




PubMed Central

A new serologic test for antibodies to Toxoplasma is described, which is based upon inhibition of specific staining with fluorescent antibody. In performing the test, a mixture of the test serum and known fluorescein-labelled antiserum is added to a dried smear of toxoplasms for 1 hour at 37°C. The smear is then rinsed and examined with a fluorescence microscope. Reduction in the brightness of fluorescence, as compared to that of a negative control slide, indicates the presence of antibody in the test serum. A comparison of the results of this test with those of the methylene blue dye test showed a strong parallelism between the two sets of results. On the other hand, the complement-fixation test for toxoplasmosis did not yield nearly as many positives as the inhibition test. The specificity of the new test was studied by comparing it with dye test results and clinical histories in human patients, and by testing a group of animals immunized with a variety of non-Toxoplasma antigens. No evidence of cross-reactions was obtained in the latter series. Some advantages and disadvantages of the inhibition test are discussed.

Goldman, Morris



Chemiluminescent lipase determination based on the enhanced luminol\\/H 2 O 2 \\/horseradish peroxidase\\/fluorescein diacetate energy transfer system  

Microsoft Academic Search

A methodology for the determination of lipase, based on the coupled processes of energy transfer and enhancement of the chemiluminescence\\u000a of the luminol-H2O2-horseradish peroxidase (HRP) system has been developed. Fluorescein diacetate (FDA) was hydrolyzed to fluorescein by the\\u000a action of the enzyme lipase, and this compound acted as an enhancer of the chemiluminescent process and acceptor of the chemiluminescent\\u000a emission

A. Navas Díaz; F. G. Sanchez; M. C. Torijas; J. Lovillo



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


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

Tabery, H M



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

PubMed Central

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

Tabery, H M



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


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

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



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


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

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



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

NASA Astrophysics Data System (ADS)

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

Kraut, Nadine D.


Technical aspects of the use of 3?,6?-diacetyl fluorescein for vital fluorescent staining of bacteria  

Microsoft Academic Search

Viable bacteria of several species have the capability to incorporate 3?,6?-diacetyl fluorescein (FDA) and rapidly hydrolyze\\u000a it to fluorescein, which is stored intracellularly. However, several strains of viableEscherichia coli andAlcaligenes faecalis do not evolve and accumulate significant amounts of fluorescein when incubated on glass slides in the presence of FDA. In\\u000a the present study, 105–107\\u000a E. coli orA. faecalis bacteria

Gustaf Brunius



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

PubMed Central

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

Hoff, K A



Fluorescein isothiocyanate staining and characterization of avian heterophils 1 Mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the USDA and does not imply its approval to the exclusion of other products that may be suitable. 1  

Microsoft Academic Search

Fluorescein isothiocyanate (FITC) was found to stain cytoplasmic granules of avian heterophil-granulocytes. In tissue sections, the fluorescent granulocytes were predominantly distributed adjacent to trabecular bones. The fluorescein stained granulocytes were abundant in synovial fluids of chickens with synovitis. A significant correlation was observed in the percent of fluorescein labeled granulocytes in blood smears and the percent of heterophils determined using

N. C Rath; G. R Huff; J. M Balog; W. E Huff



RED Facts: Chlorhexidine Diacetate.  

National Technical Information Service (NTIS)

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



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....



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....



The photography of fluorescein  

SciTech Connect

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

Welch, J.D.



Assessment of antifungal effects of a novel compound from Burkholderia cepacia against Fusarium solani by fluorescent staining  

Microsoft Academic Search

A novel compound CF66I produced by Burkholeria cepacia was investigated for its antifungal effects against Fusarium solani by three different fluorescent dyes. Dual staining with propidium iodide (PI) and fluorescein diacetate (FDA) demonstrated\\u000a high doses of CF66I (120.0 ?g ml?1) killed the fungi by acting primarily on the cell membrane. However, at fungistatic concentration (20.0 ?g ml?1) of this compound, microscopic observations revealed swelling hyphae with

Xin Li; Chun-Shan Quan; Hui-Ying Yu; Jian-Hua Wang; Sheng-Di Fan



Fluorescein Redirects a Ruthenium-Octaarginine Conjugate to the Nucleus  

PubMed Central

The cellular uptake and localization of a Ru-octaarginine conjugate with and without an appended fluorescein are compared. The inherent luminescence of the Ru(II) dipyridophenazine complex allows observation of its uptake without the addition of a fluorophore. Ru-octaarginine-fluorescein stains the cytosol, nuclei and nucleoli of HeLa cells under conditions where the Ru-octaarginine conjugate without fluorescein shows only punctate cytoplasmic labeling. At higher concentrations, however, Ru-octaarginine without the fluorescein tag does exhibit cytoplasmic, nuclear, and nucleolar staining. Attaching fluorescein to Ru-octaarginine lowers the threshold concentration required for diffuse cytoplasmic labeling and nuclear entry. Hence, the localization of the fluorophore-bound peptide cannot serve as a proxy for that of the free peptide.

Puckett, Cindy A.; Barton, Jacqueline K.



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

National Technical Information Service (NTIS)

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



[Fluorescein angiography findings in intermediate uveitis].  


The present paper describes the results of angiographic examinations of 48 eyes (29 patients) with intermediate uveitis. More than 50% of the cases displayed pathologic changes of the retinal blood vessels, such as increased fluorescein staining of the vessel walls and leakages of the retinal veins or venules, respectively. Some 20% of the eyes manifested cystoid macular edema and/or edema of the optic disk which had gone undetected by ophthalmoscopy. These findings suggest that vascular changes may play a role in the pathogenesis of intermediate uveitis. The question as to whether this disease might be caused rather by retinal vasculitis than by uveitis is discussed. PMID:3236729

Schenck, F; Böke, W



Wood stains  


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


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

Federal Register 2010, 2011, 2012, 2013

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



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

Microsoft Academic Search

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




Anthelminthic properties of mangostin and mangostin diacetate.  


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

Keiser, Jennifer; Vargas, Mireille; Winter, Rolf



SLE retinopathy: evaluation by fluorescein angiography.  

PubMed Central

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

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



Putting vital stains in context.  


While vital staining remains a cornerstone in the diagnosis of ocular disease and contact lens complications, there are many misconceptions regarding the properties of commonly used dyes by eye-care practitioners and what is and what is not corneal staining after instillation of sodium fluorescein. Similarly, the proper use and diagnostic utility of rose Bengal and lissamine green B, the other two ophthalmic dyes commonly used for assessing ocular complications, have similarly remained unclear. Due to the limitations of vital stains for definitive diagnosis, concomitant signs and symptoms in addition to a complete patient history are required. Over the past decade, there have been many reports of a type of corneal staining--often referred to as solution-induced corneal staining (SICS)--that is observed with the use of multipurpose solutions in combination with soft lenses, more specifically silicone hydrogel lenses. Some authors believe that SICS is a sign of lens/solution incompatibility; however, new research shows that SICS may be neither a measure of lens/solution biocompatibility nor 'true' corneal staining, as that observed in pathological situations. A large component of SICS may be a benign phenomenon, known as preservative-associated transient hyperfluorescence (PATH). There is a lack of correlated signs and/or symptoms with SICS/PATH. Several properties of SICS/PATH, such as appearance and duration, differentiate it from pathological corneal staining. This paper reviews the properties of vital stains, their use and limitations in assessment of the ocular surface, the aetiology of corneal staining, characteristics of SICS/PATH that differentiate it from pathological corneal staining and what the SICS/PATH phenomenon means for contact lens-wearing patients. PMID:23051047

Efron, Nathan



Photobleaching of disodium fluorescein in water  

Microsoft Academic Search

Photobleaching of disodium fluorescein dissolved in water is experimentally investigated using laser induced fluorescence\\u000a (LIF). It is demonstrated that significant photobleaching occurs on the millisecond time scale, resulting in a large decrease\\u000a in the fluorescence signal emanating from a constant concentration sample. The importance of avoiding photobleaching when\\u000a using LIF with disodium fluorescein for concentration measurements in water flow experiments

J. R. Saylor




PubMed Central

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

Zobel, C. Richard; Beer, Michael



Evaluation of the immunological specificity of fluorescein-labelled anti-human IgM conjugates  

PubMed Central

The immunological specificity of two anti-IgM fluorescein-labelled antisera was evaluated in gel diffusion, by the direct immunofluorescent technique on characterized bone marrow preparations taken from patients with myeloma and by an indirect staining method employing virus-infected cells and selected post-infection sera. The results show that neither gel diffusion nor direct immunofluorescent methods provide a reliable index of specificity for conjugates to be used in indirect procedures.

Chantler, Shireen; Haire, Margaret



Fluorescein conjugates of 9- and 10-hydroxystearic acids: synthetic strategies, photophysical characterization, and confocal microscopy applications.  


Different strategies are presented to conjugate a fluorescein moiety to 9- and 10-hydroxystearic acids (HSAs). 5-Amino-fluorescein (5-AF) was used as a starting reagent. When reacted with acyl-chloride-modified HSAs, 5-AF gave rise to stable amide derivatives with a 75% reaction yield. These products exhibited the typical steady-state and time-resolved fluorescence properties of the fluorescein chromophore with absorption at 494 nm and emission at 519 nm. Flow cytometry studies confirmed the distinct proapoptotic effect of underivatized 9-HSA on Jurkat cells and revealed a comparable ability of its amide derivative. Confocal microscopy imaging studies showed that green fluorescence could stain intracellular membranous structures. Moreover, dual-dye labeling with Mito Tracker Red, followed by colocalization analysis, revealed that HSA can move to the mitochondria. Thus, fluorescent derivatives of HSA can be used to monitor the localization of these biologically active molecules in living cells and can provide a useful tool for linking biochemical investigation with optical visualization methods. In contrast, when unmodified HSAs were used, the reaction gave monoesterified and diesterified fluorescein derivatives. These products exhibited unusual steady-state and time-resolved fluorescence properties with the excitation wavelength at 342 nm and the emission wavelength at 432 nm. It is shown that the synthesized HSA amides of fluorescein provide all of the typical photophysical and instrumental advantages of this popular dye, whereas the unusual luminescence and excitation properties of the monoester and diester of the 5-aminofluorescein would make these dyes interesting to explore as potential candidates for two photon excitation applications. PMID:15556558

Boga, Carla; Puggioli, Silvia; Gherpelli, Marica; Farruggia, Giovanna; Pagnotta, Eleonora; Masotti, Lanfranco; Neyroz, Paolo



Absolute configuration and 1H NMR characterization of rosmaridiphenol diacetate.  


The correction of patented structure 1 of rosmaridiphenol, an antioxidant isolated from rosemary, Rosmarinus officinalis, was made recently. The correct structure is proposed as 11,12-dihydroxy-8,11,13-icetexatrien-1-one (2a) based on 2D NMR data. In order to further support the structure, this work reports the single-crystal X-ray analysis, the complete (1)H NMR assignment by full spin-spin simulation, and the absolute configuration of the diacetate 2b derived via vibrational circular dicroism measurements in comparison with density functional theory calculated data. PMID:22424272

Muñoz, Marcelo A; Perez-Hernandez, Nury; Pertino, Mariano W; Schmeda-Hirschmann, Guillermo; Joseph-Nathan, Pedro



Sputum stain for mycobacteria  


Acid fast bacilli stain; AFB stain; Tuberculosis smear; TB smear ... Abnormal results show that the stain is positive for: Mycobacterium tuberculosis Mycobacterium avium-intracellular Other mycobacteria or acid-fast bacteria


Fluorescein as a contrasting agent for breast cancer  

NASA Astrophysics Data System (ADS)

The accumulation of administrated fluorescein in some breast diseases was investigated. The fluorescein concentration in about 200 macropreparations of resected mammas was measured with ratio fluorometer. The accumulation of dye in breast cancers is significantly different from that in benign diseases when determined in more than 10 hours after the dye administration. The critical value 1.75 of fluorescein accumulation factor sets the false positive error as well as false negative error to be equal to 6.6% for the fluorescent method of diagnostics. These data may be used both in express intra-surgical fluorescent diagnostics and in NMR imaging with chemically modified fluorescein.

Lazarev, V. V.; Polsachev, V. I.



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



Fluorescein and indocyanine green angiographic findings in progressive hemifacial atrophy.  


A 20-year-old man who had progressive hemifacial atrophy was examined using fluorescein and indocyanine green angiography. The fundus showed sectional chorioretinal atrophy. Fluorescein angiography showed window defects without leakage. Indocyanine green angiography revealed narrow choroidal vessels with hypofluorescence. PMID:19213282

Kawazoe, Mariko; Hirata, Akira; Okinami, Satoshi


Differential staining of bacteria: flagella stain.  


Bacterial flagella are appendages used for motility. Their presence is a useful tool for identification and differentiation of prokaryotes. Since flagella are too thin to be seen by compound light microscopy, staining methods employ the use of a mordant (often tannic acid) to make them thick enough to see using an oil immersion objective. Two protocols are described. Basic Protocol 1 is a modified Leifson method and is the one that many microbiologists have adapted. Basic Protocol 2 is a wet-mount stain using a Ryu stain and is included because the stain is stable at room temperature. Both of these methods are fairly time-consuming, taking from 15 to as long as 60 min to perform. PMID:19885934

Breakwell, Donald P; Moyes, Rita B; Reynolds, Jackie



Congo red vital staining of cornea and conjunctiva.  


Vital staining with an aqueous solution of 1% Congo red has been studied in the slit lamp. In 98 cases the dye was mixed with 1% lissamine green, in 120 eyes subsequent staining was performed with 0.125% fluorescein, and in 80 cases the mucous thread from the inferior conjunctival fornix was microscoped. Congo red stains dead cells, degenerate cells, and mucus. The dye discloses keratitis, corneal erosion, contact lens damages, corrosions, etc. It stains like lissamine green and rose bengal, though less frequently and less intensely than these. Congo red is a pH indicator. Acid reaction beyond its pH-range (3.0-5.2) has not been demonstrated. Amyloid-specific colour reaction (red-green dichromatic polarisation) has been noticed in mucous fibrils, most often in relation to infectious conjunctivitis and corrosion, never in normal eyes. The phenomenon is believed to indicate degeneration of the mucous fibrils (on the analogy of toluidine-blue-stained mucus), whereas not presence of genuine amyloid. It is, in other words, an important phenomenon in the differential diagnosis. Congo red is hardly indicated in ordinary clinical practice for vital staining of cornea and conjunctiva. Fluorescein, combined with rose bengal or lissamin green should be preferred. PMID:62486

Norn, M S



Acid-fast stain  


The acid-fast stain is a laboratory test that determines if a sample of tissue, blood, or other body ... lab team member washes the slide with an acid solution and applies a different stain. The bacteria ...


Sputum Gram stain  


... called a smear. Stains are placed on the sample. The lab team member looks at the stained slide under a microscope, checking for bacteria and white blood cells. The color, size, and shape of the cells help identify the ...


Fluorescent staining of gels.  


Certain transition metal complexes show intensive fluorescence when bound to proteins. They can be used to stain gels after electrophoresis with a sensitivity approaching that of silver staining, but in a much simpler and more reproducible procedure. Stains can be prepared easily and at a fraction of the cost of commercially available reagents.Hydrophobic dyes can be used to stain gels without fixing; they do not interfere with later blotting or electro-elution. PMID:22585519

Buxbaum, Engelbert



Port-Wine Stain  


newsletter | contact Share | Port-Wine Stain A parent's guide for infants and babies A A A This is a three-month-old infant with a port-wine stain. Overview A port-wine stain is a type of birthmark that is caused by a ...


Ultra-widefield fluorescein angiography of white without pressure  

PubMed Central

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

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



Anterior capsule staining for capsulorhexis in cases of white cataract  

Microsoft Academic Search

Purpose: To compare the safety and efficacy of trypan blue 0.1%, gentian violet 0.001%, indocyanine green 0.5% (ICG), fluorescein 2%, and the patient's autologous blood for anterior capsule staining in cases of white cataract.Setting: Rajendra Prasad Center for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India.Methods: Fifty eyes of 50 patients with age-related white cataract had anterior

Vijay K Dada; Namrata Sharma; Rajeev Sudan; Harinder Sethi; Tanuj Dada; Mayank S Pangtey



Causes of Bridge Pier Staining.  

National Technical Information Service (NTIS)

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

S. I. Thornton C. Springer



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

SciTech Connect

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

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



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



Binding and internalization of ricin labelled with fluorescein isothiocyanate.  


The toxic lectin ricin has been covalently labelled with fluorescein isothiocyanate on the enzymatically active A chain. The fluorescein reacted toxin maintains its biological activity. The lateral diffusion coefficient of cell surface bound ricin, studied in two cell lines by fluorescence photobleaching recovery, is D = 1 - 2 x 10(-10) cm2/s. Fluorescence microscopy provides preliminary evidence for secondary endosomes in the cytoplasm. PMID:2113381

Bellelli, A; Ippoliti, R; Brunori, M; Kam, Z; Benveniste, M; Emmanuel, F; Turpin, E; Alfsen, A; Frénoy, J P



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


Here we demonstrate that the semi-synthetic flavonoid ayanin diacetate induces cell death selectively in leukemia cells without affecting the proliferation of normal lymphocytes. Incubation of human leukemia cells with ayanin diacetate induced G(2)-M phase cell cycle arrest and apoptosis which was prevented by the non-specific caspase inhibitor z-VAD-fmk and reduced by the overexpression of Bcl-x(L). Ayanin diacetate-induced cell death was found to be associated with: (i) loss of inner mitochondrial membrane potential, (ii) the release of cytochrome c, (iii) the activation of multiple caspases, (iv) cleavage of poly(ADP-ribose) polymerase and (v) the up-regulation of death receptors for TRAIL, DR4 and DR5. Moreover, the combined treatment with ayanin diacetate and TRAIL amplified cell death, compared to single treatments. These results provide a basis for further exploring the potential applications of this combination for the treatment of cancer. PMID:23063980

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



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

PubMed Central

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

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



Gamma irradiation of fine-emulsion sausage containing sodium diacetate.  


Listeria monocytogenes, a psychrotrophic foodborne pathogen, is a frequent postprocess contaminant of ready-to-eat (RTE) meat products, including frankfurters and bologna. Ionizing radiation can eliminate L. monocytogenes from RTE meats. Sodium diacetate (SDA) incorporated into fine-emulsion sausages inhibits the growth of L. monocytogenes. Irradiation of L. monocytogenes suspended in SDA solutions resulted in synergistic reductions of the microorganism. L. monocytogenes populations were reduced by > 9 log10 units at a radiation dose of 1.5 kGy when suspended in 0.125% SDA solution. In contrast, the D10-values (the ionizing radiation doses required to reduce the population by 90%) were 0.58, 0.59, 0.57, and 0.53 kGy for L. monocytogenes populations suspended in emulsions containing 0, 0.125, 0.25, and 0.5% SDA, respectively. The D10-values for L. monocytogenes surface inoculated onto frankfurters dipped in 0, 0.125, 0.25, and 0.5% SDA solutions were 0.58, 0.53, 0.54, and 0.52 kGy, respectively. Postirradiation growth of L. monocytogenes suspended in beef bologna emulsion at 9 degrees C was dependent on SDA concentration and ionizing radiation dose. Very small, but statistically significant, changes in bologna redness, lipid oxidation, and shear force were observed for the beef bologna emulsion with the highest SDA concentration (0.5%) and irradiation dose (3.0 kGy). SDA can inhibit the proliferation of L. monocytogenes surviving the irradiation process with minimal impact on fine-emulsion sausage color, lipid oxidation, and firmness when used within regulatory limits. PMID:12747691

Sommers, Christopher; Fan, Xuetong



Gram stain of tissue biopsy  


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


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

SciTech Connect

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

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



Fluorescein angiographic defects of the optic disc in glaucoma.  


An improved technique for high-contast, high-resolution fluorescein angiography of the optic disc has been developed that almost eliminates pseudofluorescence. Fluorescein angiography of the optic disc was performed on normal, ocular hypertensive, and glaucomatous patients. Rapid-sequence angiograms in the early arterial phases have demonstrated localized areas of hypofluorescence or filling defects of the optic disc. Two types of fluorescein filling defects were observed-absolute and relative. The number of absolute filling defects, which increased with degree of visual field loss, was greater in glaucomatous than in ocular hypertensive or normal eyes. Similarly, ocular hypertensive eyes showed a larger number of discs with filling defects than normal eyes. It is postulated that relative defects progress to absolute filling defects, which may be an indication of impending loss of visual field. PMID:921573

Schwartz, B; Rieser, J C; Fishbein, S L



Experimental contamination of Minims of fluorescein by Pseudomonas aeruginosa.  

PubMed Central

Contamination of fluorescein solutions by Pseudomonas aeruginosa has been a concern of ophthalmologists for many years because of the severity of pseudomonas keratitis. Attempts to prevent contamination have been directed at stringent sterility control during manufacture and the introduction of single-dose disposable containers such as Minims. Deliberate contamination of Minims fluorescein with Pseudomonas aeruginosa was attempted. Under conditions likely to be met with in clinical practice the contents remained sterile. However, under extreme conditions of immersion in pure broth culture of Pseudomonas aeruginosa contamination could be achieved. The relevance of these results to clinical practice is discussed.

Claoue, C



Advances in modifying fluorescein and rhodamine fluorophores as fluorescent chemosensors.  


The fluorophores based on xanthene scaffolds, mainly containing rhodamine and fluorescein dyes, have attracted considerable interest from chemists due to their excellent photophysical properties such as high absorption coefficient, high fluorescence quantum yield, high photostability and relatively long wavelengths of fluorescence emission spectra. In this feature article, we overview the strategies in the development of fluorescent probes that are operating through the modification of the skeletons of fluorescein and rhodamine dyes, and the fluorescent behaviors of these probes toward specific analyte are discussed. PMID:23164947

Zheng, Hong; Zhan, Xin-Qi; Bian, Qing-Na; Zhang, Xiao-Juan



Toxoplasmosis I. Studies by the Fluorescein-labelled Antibody Technique  

PubMed Central

The fluorescein-labelled antibody technique was investigated for the diagnosis of toxoplasmosis. The direct method, the inhibition and indirect modifications are suitable for the demonstration of Toxoplasma gondii in fluid and tissue-impression slides from animals in the acute phase of infection. The method was not applicable with the frozen tissue sections. The fluorescein-labelled antibody inhibition technique detected antibodies in immune sera from various species of animal. However the titres obtained were lower than with the complement-fixation test. ImagesFigure I

Ruckerbauer, G. M.; Robertson, A.; Bannister, G. L.; Boulanger, P.; Beauregard, M.



Joint fluid Gram stain  


... and shape of the cells help identify the bacteria. ... Abnormal results mean bacteria were seen on the Gram stain. This may be a sign of a joint infection, for example, gonococcal arthritis or arthritis due to Staphylococcus aureus.


Candida, fluorescent stain (image)  


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


Kinetics of staining and bleaching  

Microsoft Academic Search

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

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



Electronic spectroscopy of biomolecules in solution: fluorescein dianion in water  

Microsoft Academic Search

A combined and sequential Monte Carlo–quantum mechanics methodology is used to describe the electronic absorption spectrum of the fluorescein dianion in water. Different sets of 100 statistically relevant configurations composed of the solute and several solvent molecules are sampled from the Monte Carlo simulation for a posteriori quantum mechanical calculations of the spectra. In the largest case the configurations are

Daniel L. Silva; Kaline Coutinho; Sylvio Canuto



Endochitinase activity determination using N-fluorescein-labeled chitin  

Microsoft Academic Search

A fluorimetric method for the determination of endochitinolytic activity using N-fluorescein-labeled chitin (FITC-Chitin) is proposed, and a procedure for FITC-Chitin preparation with a degree of FITC content of 2.2 mol% (one FITC molecule per 45 glucosamine residues) is described. FITC-Chitin is capable to distinguish endochitinase and exochitinase (?-N-acetylglucosaminidase) activities.

Vladimir E Tikhonov; Luis V Lopez-Llorca; Jesús Salinas; Elena Monfort



Nonlinear Optical Phase Conjugation in Fluorescein and Bismuth Silicate (BSO).  

National Technical Information Service (NTIS)

The goal of the experiment was to produce nonlinear optical phase conjugation (NOPC) in fluorescein-doped boric acid glass and in BSO, and to use two-wave mixing (TWM) in BSO to transfer energy between laser beams. Next, the parameters that affect these p...

P. R. Leatherman



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.



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

PubMed Central

Evolution of reactive oxygen species (ROS), generated during the patho-physiological stress of nervous tissue, has been implicated in the etiology of several progressive human neurological disorders including Alzheimer’s disease (AD) and amylotrophic lateral sclerosis (ALS). In this brief communication we used mixed isomers of 5-(and-6)-carboxy-2?,7?-dichlorofluorescein diacetate (carboxy-DCFDA; C25H14Cl2O9; MW 529.3), a novel fluorescent indicator, to assess ROS generation within human neuronal-glial (HNG) cells in primary co-culture. We introduced pathological stress using the sulfates of 12 environmentally-, industrially- and agriculturally-relevant divalent and trivalent metals including Al, Cd, Cu, Fe, Hg, Ga, Mg, Mn, Ni, Pb, Sn and Zn. In this experimental test system, of all the metal sulfates analyzed, aluminum sulfate showed by far the greatest ability to induce intracellular ROS. These studies indicate the utility of using isomeric mixtures of carboxy-H2DCFDA diacetates as novel and highly sensitive, long-lasting, cell-permeant, fluorescein-based tracers for quantifying ROS generation in intact, metabolizing human brain cells, and in analyzing the potential epigenetic contribution of different metal sulfates to ROS-generation and ROS-mediated neurological dysfunction.

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



Shimmering Stained Glass.  

ERIC Educational Resources Information Center

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

Simon, Gail Murray



"Stained Glass" Landscape Windows  

ERIC Educational Resources Information Center

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

Vannata, Janine



Lineage Labeling of Zebrafish Cells with Laser Uncagable Fluorescein Dextran  

PubMed Central

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 labeling1 and pressure injection of traceable enzymes2 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 time3-5. 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 zebrafish6-8. The optical clarity of the zebrafish embryo and the relative ease of transgenesis have made these particularily attractive tools for lineage labeling and to observe the migration of cells in vivo7. 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 fluorescence9. 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 pinhole10. In addition, antibodies against fluorescein detect only the uncaged form, and the epitope survives fixation well11. Finally, caged fluorescein can be activated with very high 3-D resolution, especially if two-photon microscopy is employed 12,13. This protocol describes a method of lineage labeling by caged fluorescein and laser uncaging. Subsequenctly, uncaged fluorescein is detected simultaneously with other epitopes such as GFP by labeling with antibodies.

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



Lineage labeling of zebrafish cells with laser uncagable fluorescein dextran.  


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



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...


Effect of Fluorocarbon Surfactant Additives on the Effective Viscosity of Acetone Solutions of Cellulose Diacetate.  

National Technical Information Service (NTIS)

The fact that the viscosity of acetone solutions of cellulose diacetate (CDA) DATs can be reduced by the addition of small quantities of water has been known for a long time. It is given practical application in the production of acetone fiber. An analogo...

L. A. Shits N. Y. Kal'nova



Synthesis and antioxidant activity of a novel class of fluorescein-based ?-C-glycosides.  


A series of fluorescein-based ?-C-glycosyl ketones were synthesized through aldol condensation of ?-C-glycosyl ketones with fluorescein monoaldehyde under ambient reaction conditions in good yields. Formation of the expected product has been confirmed through different spectral techniques. Fluorescein-based ?-C-glycosides show moderate anti-oxidant activities with maximum inhibitory activity of 60%. PMID:23867296

Rajasekar, Mani; Das, Thangamuthu Mohan



Stain length passive dosimeters  

SciTech Connect

Passive dosimeters with instant readout capability have been devised by combining the principles of a gas indicator tube with membrane control of mass transfer. The membrane controls the diffusion of the gas or vapour to the reagent-impregnated support where it reacts to produce a stain. Results with H/sub 2/S and benzene monitors demonstrate that time-weighted average concentration of ambient gas or vapour can be measured accurately and precisely by following the movement of the coloured stain in the specially prepared and calibrated indicator tube. The 95% confidence interval of such measurements at the ThV is +/-20% for H/sub 2/S and +/-15% for benzene, well within NI0SH limits of acceptability.

Sefton, M.V.



Stain length passive dosimeters  

SciTech Connect

Passive dosimeters with instant readout capability have been devised by combining the principles of a gas indicator tube with membrane control of mass transfer. The membrane controls the diffusion of the gas or vapor to the reagent impregnanted support where it reacts to produce a stain. Results with H/sub 2/S and benzene monitors demonstrated that time weighted average concentration of ambient gas or vapor can be measured accurately and precisely by following the movement of the colored stain in the specially prepared and calibrated indicator tube. The 95% confidence interval of such measurements at the TLV (80 ppm-hrs) is +/- 20% for H/sub 2/S and +/- 15% for benzene, well within NIOSH limits of acceptability.

Sefton, M.V.; Kostas, A.V.; Lombardi, C.



Fluoresceinated phosphoethanolamine for flow-cytometric measurement of lipid peroxidation.  


A new lipophilic fluorescein probe (fluor-DHPE) has been identified that can assay lipid peroxidation in mammalian cells on a cell-by-cell or selected-cell-subpopulation basis by flow cytometry. Application of this approach requires that the fluorescent probe be nonexchangeable among cells. Fluorescein is an appropriate fluorophore, since its fluorescence matches the specifications of common flow cytometers and the compound loses its fluorescence upon reaction with peroxyl radicals. Upon examination of four lipophilic derivatives of fluorescein, fluor-DHPE was found to be the only probe that was nonexchangeable among labeled and unlabeled rat RBC for at least 24 h. The exposure of fluor-DHPE-labeled RBC to benzoyl peroxide followed by mixing the sample with RBC unexposed to peroxide led to a decrease in fluorescence. Furthermore, the flow cytometer could clearly select the subpopulation of cells undergoing lipid peroxidation from those cells that were not. Fluor-DHPE-labeled-RBC obtained from rats and exposed to cumene hydroperoxide also displayed a gradual decrease in fluorescence. This decrease was preventable by either regulation of the vitamin E content in the animal diet or in vitro supplementation of cells with vitamin E. We conclude that fluor-DHPE is a stable and nonexchangeable probe for monitoring lipid peroxidation in cell subpopulations by flow cytometry. PMID:9801063

Maulik, G; Kassis, A I; Savvides, P; Makrigiorgos, G M



Blood stain pattern analysis  

Microsoft Academic Search

Bloodstain pattern analysis (BPA) refers to the collection, categorization and interpretation of the shape and distribution\\u000a of bloodstains connected with a crime. These kinds of stains occur in a considerable proportion of homicide cases. They offer\\u000a extensive information and are an important part of a functional, medically and scientifically based reconstruction of a crime.\\u000a The following groups of patterns can

O. Peschel; S. N. Kunz; M. A. Rothschild; E. Mützel



Increased NO bioavailability in aging male rats by genistein and exercise training: using 4, 5-diaminofluorescein diacetate  

PubMed Central

Background Several kinds of anti-oxidants have drawn a lot of intension for their benefits on vascular protection. In addition, it has been demonstrated that exercise training could improve endothelial function by up-regulating endothelial nitric oxide synthase (eNOS) protein. Therefore, the present study aims to investigate the effects of genistein, a potent phyto-antioxidant, and exercise training on age-induced endothelial dysfunction in relation to NO bioavailability using in situ NO-sensitive fluorescent dye detection. Methods Male Wistar rats (20-22-month old) were divided into four groups: aged rats treated with corn oil, (Aged+Veh, n = 5), aged rats treated with genistein (Aged+Gen, n = 5, (0.25 mg/kg BW/day, s.c.)), aged rats with and without exercise training (Aged+Ex, n = 5, swimming 40 min/day, 5 days/week for 8 weeks) (Aged+Without-Ex, n = 5). Cremaster arterioles (15-35 micrometer) were visualized by fluorescein isothiocyanate labeled dextran (5 microgram/ml). The vascular response to acetylcholine (Ach; 10-5M, 5 ml/5 min) was accessed after 1-min norepinephrine preconstriction (10 micro molar). To determine NO bioavailability, the Krebs-Ringer buffer with 4, 5-diaminofluorescein-diacetate (3 micro molar DAF-2DA), and 10 micro- molar Ach saturated with 95%N2 and 5%CO2 were used. Changes of DAF-2T-intensities along the cremaster arterioles were analyzed by the Image Pro-Plus Software (Media Cybernatics, Inc, USA). Liver malondialdehyde (MDA) level was measured by thiobarbituric acid reaction and used as an indicator for oxidative stress. Results The results showed that means arterial blood pressure for both Aged+Gen and Aged+Ex groups were significantly reduced when compared to the Aged groups, Aged+Veh and Aged+Without-Ex (P < 0.05). Among the treated groups, Ach-induced vasodilatation were significantly increased (P < 0.05) and was associated with increased NO-associated fluorescent intensities (P < 0.05). On the other hand, MDA levels were significantly reduced (P < 0.05) when Aged+Veh was compared to Aged+Without-Ex. Conclusion These findings showed that genistein and exercise training could improve age-induced endothelial dysfunction and is related to the increased NO bioavailability.

Eksakulkla, Sukanya; Suksom, Daroonwan; Siriviriyakul, Prasong; Patumraj, Suthiluk



Coffee stain technique.  


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

Hodge, Gerald P



Blood stain pattern analysis.  


Bloodstain pattern analysis (BPA) refers to the collection, categorization and interpretation of the shape and distribution of bloodstains connected with a crime. These kinds of stains occur in a considerable proportion of homicide cases. They offer extensive information and are an important part of a functional, medically and scientifically based reconstruction of a crime. The following groups of patterns can essentially be distinguished: dripped and splashed blood, projected blood, impact patterns, cast-off stains, expirated and transferred bloodstains. A highly qualified analysis can help to estimate facts concerning the location, quality and intensity of an external force. A sequence of events may be recognized, and detailed questions connected with the reconstruction of the crime might be answered. In some cases, BPA helps to distinguish between accident, homicide and suicide or to identify bloodstains originating from a perpetrator. BPA is based on systematic training, a visit to the crime scene or alternatively good photographic documentation, and an understanding and knowledge of autopsy findings or statements made by the perpetrator and/or victim. A BPA working group has been established within the German Society of Legal Medicine aiming to put the knowledge and practical applications of this subdiscipline of forensic science on a wider basis. PMID:21069481

Peschel, O; Kunz, S N; Rothschild, M A; Mützel, E



Cellular Uptake Behavior of Fluorescein: Intercalated Layered Double Hydroxide  

NASA Astrophysics Data System (ADS)

In order to define the ability of layered double hydroxide (LDH) as materials for drug delivery, fluorescein (Fluo) anion intercalated LDH (Fluo/LDH) was synthesized by hydrothermal treatment and observed the cellular uptake of the Fluo/LDH for mammalian cell (L929). The synthesized Fluo/LDH showed a LDH structure, high fluorescence and low cytotoxicity. According to the fluorescence, confocal and TEM images of cells, the Fluo/LDH seemed to be internalized into the L929 cell by cellular endocytosis and dissolved inside the cell to exhibit the fluorescence of cellular cytoplasm.

Tanaka, Miyuki; Aisawa, Sumio; Hirahara, Hidetoshi; Narita, Eiichi; Yin, Shu; Sato, Tsugio



Synthesis, purification and kinetic properties of fluorescein-labelled penicillins.  

PubMed Central

The synthesis and properties of six fluorescein-labelled penicillins are reported. The two isomers of fluoresceyl-glycyl-6-amino-penicillanic acid are probably the best compounds to use for detection of all the penicillin-binding proteins (PBPs) present in a bacterial membrane preparation. However, the derivatives of ampicillin were much more efficient against Enterobacter aerogenes PBP3. The two isomers obtained when a commercial mixture of the two isomers of carboxyfluorescein was used most often exhibited similar properties, but the Streptomyces R61 extracellular DD-peptidase was only efficiently acylated by the 5'-carboxyfluorescein derivative of glycyl-6-aminopenicillanic acid.

Lakaye, B; Damblon, C; Jamin, M; Galleni, M; Lepage, S; Joris, B; Marchand-Brynaert, J; Frydrych, C; Frere, J M



Kinetics of the polymerization of methylhydroquinone diacetate, terephthalic acid and poly(ethylene-terephthalate)  

Microsoft Academic Search

Poly(ethylene terephthalate) (PET) was modified using methyl hydroquinone diacetate (MHQDA) and terephthalic acid (TA) to develop copolyester membranes for pervaporation separation. These membranes are expected to have improved thermomechanical properties. Two different polymer compositions (PET 70\\/30 (MHQDA+TA) and PET 50\\/50 (MHQDA+TA)) were synthesized using the melt polymerization route. The melt polymerization kinetics of these systems are reported. Two different polyesterification

Habib I Shaban; Hamad Al-Adwani; Anaam Behbehani; Johnson Mathew



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


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



Organic Reactions in Ionic Liquids: Gewald Synthesis of 2?Aminothiophenes Catalyzed by Ethylenediammonium Diacetate  

Microsoft Academic Search

Ionic liquids based on 1?butyl?3?methylimidazolium tetrafluoroborate (BmimBF4) and 1?butyl?3?methylimidazolium hexafluorophosphate (BmimPF6) were used as reusable alternatives to volatile organic solvents (VOCs) for ethylenediammonium diacetate (EDDA) catalyzed Gewald synthesis of 2?aminothiophenes. Significant rate enhancement and improvement of the yield were observed. The ionic liquids containing catalyst EDDA were recycled several times with no decreases in yields and reaction rates.

Yi Hu



Fragrance material review on 1,2-ethanediol, 1-phenyl-, 1,2-diacetate.  


A toxicologic and dermatologic review of 1,2-ethanediol, 1-phenyl-, 1,2-diacetate when used as a fragrance ingredient is presented. 1,2-Ethanediol, 1-phenyl-, 1,2-diacetate is a member of the fragrance structural group Aryl Alkyl Alcohol Simple Acid Esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for 1,2-ethanediol, 1-phenyl-, 1,2-diacetate were evaluated, then summarized, and includes physical properties data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE fragrances. PMID:22406575

McGinty, D; Letizia, C S; Api, A M



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

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



Does fundus fluorescein angiography procedure affect ocular pulse amplitude?  


Purpose. This study examines the effects of fundus fluorescein angiography (FFA) procedure on ocular pulse amplitude (OPA) and intraocular pressure (IOP). Materials and Methods. Sixty eyes of 30 nonproliferative diabetic retinopathy patients (15 males, 15 females) were included in this cross-sectional case series. IOP and OPA were measured with the Pascal dynamic contour tonometer before and after 5 minutes of intravenous fluorescein dye injection. Results. Pre-FFA mean OPA value was 3.05 ± 1.36?mmHg and post-FFA mean OPA value was 2.93 ± 1.28?mmHg (P = 0.071). Pre-FFA mean IOP value was 17.97 ± 1.99?mmHg and post-FFA mean IOP value was 17.81 ± 2.22?mmHg (P = 0.407). Conclusion. Although both mean OPA and IOP values were decreased after FFA procedure, the difference was not statistically significant. This clinical trial is registered with Australian New Zealand Clinical Trials Registry number ACTRN12613000433707. PMID:23984045

Pekel, Gökhan; Acer, Semra; Yagci, Ramazan; Cetin, Ebru Nevin; Hiraali, Mehmet Can; Kaya, Hüseyin



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

SciTech Connect

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

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



Fluorescein conjugates of 9- and 10-hydroxystearic acids: synthetic strategies, photophysical characterization, and confocal microscopy applications  

Microsoft Academic Search

Different strategies are presented to conjugate a fluorescein moiety to 9- and 10-hydroxystearic acids (HSAs). 5-Amino-fluorescein (5-AF) was used as a starting reagent. When reacted with acyl-chloride-modified HSAs, 5-AF gave rise to stable amide derivatives with a 75% reaction yield. These products exhibited the typical steady-state and time-resolved fluorescence properties of the fluorescein chromophore with absorption at 494nm and emission

Carla Boga; Silvia Puggioli; Marica Gherpelli; Giovanna Farruggia; Eleonora Pagnotta; Lanfranco Masotti; Paolo Neyroz



Pdr12p-dependent and -independent fluorescein extrusion from baker's yeast cells  

Microsoft Academic Search

Fluorescein efflux from S. cerevisiae cells was measured to study the peculiarities of fluorescein transport system, which is important for yeast resistance to certain drugs and weak organic acid preservatives. Glucose-independent and glucose-stimulated fluorescein effluxes were character - ized using iodoacetate, cyanide and orthovanadate, inhibitors of glycolysis, electron transport chain, and ATPases, respectively. It is supposed that in glucose-free medium

Volodymyr Lushchak; Oleksandra Abrat; Halyna Semchyshyn


Fluorescein mercury(II) acetate and sodium fluorescein as reagents for the determination of bis(2-chloroethyl)sulfide by fluorescence quenching  

Microsoft Academic Search

Bis(2-chloroethyl)sulfide causes strong quenching of sodium fluorescein and fluorescein mercury(II) acetate fluorescence in alcoholic media. Sodium fluorescein shows the greater promise as a reagent for its determination by fluorescence spectrometry with excitation at 366nm and measurement at 518nm. The results indicate a working range between about 3×10?7 and 4.5×10?5moll?1 bis(2-chloroethyl)sulfide in propan-l-ol. Reproducibility of the quenching effect was around ±10%

Anya L. Hunt; John F. Alder



Deformable registration of retinal fluorescein angiogram sequences using vasculature structures.  


State-of-the-art deformable registration algorithms do not perform as well with FA sequences because they are designed to deal with changes of content appearance (e.g., due to different sensors imaging the same organs) but not with content changes, which occur throughout a FA sequence as different portions or the vascular structure are visible (perfused) in different frames. This paper presents a frame-to-frame registration algorithm for ultra-wide-field-of-view (UWFV) fluorescein angiograms (FA) of the retina, based on deformable alignment of the retinal vasculature structure. Comparative experiments on an initial set of UWFV FAs indicate that, thanks to its specialization, our technique outperforms one of the best state-of-the-art methods for multimodal image registration when dealing with the demanding characteristics of the UWFV FA sequences. PMID:21096457

Perez-Rovira, Adria; Trucco, Emanuele; Wilson, Peter; Liu, Jiang



Fluorescein angiography using ultra-high speed film.  


We used a new ultra-high speed black-and-white film, Kodak T-MAX P3200 (ASA 3200), in routine fluorescein angiography on 51 patients and one normal volunteer. The increased film speed permitted a lower flash intensity than with other available films. Photophobia was improved subjectively in 25% of patients who had undergone prior angiography at a higher flash setting. In addition, patient cooperation was improved as evidenced by a decrease in the frequency of photographic artifacts and uninterpretable photographs compared with prior angiograms done using Kodak TRI-X Pan film (ASA 400). T-MAX yielded excellent resolution, but its grain size was slightly greater than that of TRI-X Pan. The use of ultra-high speed film and reduced light intensity may benefit the patient and improve photographic quality in some individuals with photophobia. PMID:2221708

Choromokos, E A; Wilson, C A; Raymond, L A; Lipman, M J



Fluorescent staining of resting spores of Polymyxa betae as a fungal vector of rhizomania disease of sugar beet in soil  

Microsoft Academic Search

When the resting spores of Polymyxa betae were pretreated with 2% sodium dodecyl sulfate (SDS) and then stained with various fluorochromes including 3,3?-dihexyloxacarbocyanine\\u000a iodide [DiOC6(3)], calcofluor, and a fluorescein isothiocyanate (FITC)-conjugated lectin, such as wheat germ lectin (WGA)\\u000a or caster bean lectin, most spores fluoresced brightly. FITC-WGA mainly stained the cell surface, while DiOC6(3) stained the\\u000a cytoplasm. After pretreatment with

Mitsuru Sayama; Yoji Momota; Shigehito Takenaka



An evaluation of some commerical Romanowsky stains  

Microsoft Academic Search

The staining properties of 43 commerical Romanowsky-type stains have been studied. Considerable differences in the appearance of stained blood films were observed with different batches of these stains, the staining of red cells being particularly variable. Attempts have been made to correlate staining patterns with stain composition as revealed by thin-layer chromatography and sulphated ash analyses. In this way it

P N Marshall; S A Bentley; S M Lewis



The Use of Fluorescent Antibody Staining in the Diagnosis of Rabies  

PubMed Central

Brain material from 750 domestic and wild animals submitted to this laboratory for rabies diagnosis was studied by the following three methods: a) microscopic examination of Williams' stained impressions, b) mouse inoculation test, and c) microscopic examination of impressions stained with fluorescein-tagged antibodies. The purpose of this investigation was to compare the sensitivity of the fluorescent antibody technique with that of two classical methods, when applied to the routine diagnosis of rabies. From the results obtained by one or the other method of study, 175 specimens were diagnosed as positive. Of these, only 58 (33 per cent) were detected by the examination of Williams' stained impressions. On the other hand, two rabid cases were missed by the mouse inoculation test, and four by the fluorescent antibody technique. Without being completely reliable, the last two methods proved to be almost equally sensitive and much more so than the examination of Williams' stained impressions. ImagesFigure 1.Fig. 2.Fig. 3.

Beauregard, M.; Boulanger, P.; Webster, W. A.



Chromatin staining of Drosophila testes.  


This protocol describes chromatin staining of Drosophila testes. To visualize DNA, preparations fixed using methanol-acetone, paraformaldehyde, or formaldehyde can be stained with several DNA-binding dyes. If the slides are to be examined with a fluorescence microscope equipped with filters that permit ultraviolet (UV) excitation, suitable dyes for DNA staining are Hoechst 33258 or 4',6-diamidino-2-phenylindole (DAPI). If the slides are to be analyzed with a confocal microscope not equipped with a UV laser, DNA can be stained with either propidium iodide or TOTO-3 iodide. PMID:22854562

Bonaccorsi, Silvia; Giansanti, Maria G; Cenci, Giovanni; Gatti, Maurizio



Emission quenching via intramolecular electron transfer for fluorescein conjugates. Dependences on driving force and medium  

Microsoft Academic Search

The fluorescence properties of conjugates of the dye, fluorescein, in which triazine or dinitrobenzoyl groups are attached to an amine function in the 5? position have been studied. The substantial quenching of fluorescence which is observed with this modification is attributed to intramolecular electron transfer involving the xanthene moiety as electron donor (i.e., the shift of anionic charge). The fluorescein

Guilford Jones; Xiaohua Qian



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

Microsoft Academic Search

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

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



Measurement of blood flow in hypospadias flaps with subvisual doses of fluorescein.  


Hypospadias flap viability is determined most often by direct observation or injection of visual doses of fluorescein and observation with a Woods lamp. We support the use of a dermofluorometer and subvisual doses of fluorescein to measure tissue fluorescence in predicting flap viability. PMID:3723677

Walker, R D; Graham, B



Field's stain – a rapid staining method for Acanthamoeba spp  

Microsoft Academic Search

Acanthamoeba sp. is a free-living amoeba known to cause chronic central nervous system infection or eye infection in humans. Many cases\\u000a remain undetected for want of a good detection system. We report for the first time a rapid staining method to facilitate\\u000a the identification of Acanthamoeba sp. using the modified Field's staining technique. A. castellanii, which was used in the

M. Pirehma; K. Suresh; S. Sivanandam; A. Khairul Anuar; K. Ramakrishnan; G. Suresh Kumar



Differential nucleolar staining affinity with a modified Papanicolaou staining procedure.  


A modified Papanicolaou staining procedure using diluted Harris' hematoxylin with potassium alum is described. Nucleolar staining varies from blue to bright red. This technique has been applied to mammary tumor cell lines in vitro under several conditions of hormonal stimulation known to induce protein synthesis and cell differentiation. Blue nucleoli are observed in control resting cells, while bright red nucleoli are seen after hormonal stimulation. PMID:6171053

Mouriquand, J; Mouriquand, C; Petitpas, E; Louis, J; Mermet, M A



Systematic study of fluorescein-functionalized macrophotoinitiators for colorimetric bioassays.  


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



Ultrawide-field Fluorescein Angiography for Evaluation of Diabetic Retinopathy  

PubMed Central

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

Kong, Mingui; Lee, Mee Yon



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.

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.



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

PubMed Central

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

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



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;



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


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

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



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


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

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



Bioavailability and pharmacokinetics of norethisterone in women after oral doses of ethynodiol diacetate.  


Measurement by radioimmunoassay of plasma norethisterone (NE) has been used to compare the bioavailability of tablets containing ethynodiol diacetate (EDA) with that of a standard oral solution of this progestogen in 12 normal women. The tablets investigated were from three batches which showed different in vitro dissolution rates. There were no significant differences in the bioavailability of the tablet formulations, which were essentially bioequivalent to the solution. Peak blood levels of NE were reached within 4h of EDA administration in solution or tablets. After the peak, NE plasma levels declined in two phases, with a mean terminal elimination half lives of 4 to 6.9h. The pharmacokinetics of NE after EDA administration showed some similarity to those observed by other workers after oral doses of NE itself. PMID:428229

Vose, C W; Butler, J K; Williams, B M; Stafford, J E; Shelton, J R; Rose, D A; Palmer, R F; Breckenridge, A M; Orme, M L; Serlin, M J




Technology Transfer Automated Retrieval System (TEKTRAN)

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


Rapid assembly of oligosaccharides: 1,2-diacetal-mediated reactivity tuning in the coupling of glycosyl fluorides  

Microsoft Academic Search

This paper describes the application of 1,2-diacetal protecting groups to control the reactivity tuning of glycosyl fluorides in oligosaccharide coupling reactions. The synthetic potential of this new methodology is demonstrated by the ‘one-pot’ synthesis of a linear pentasaccharide and the efficient assembly of the core oligosaccharide of the GPI anchor of yeast (Saccharomyces cerevisiae).

Daniel K Baeschlin; Luke G Green; Michael G Hahn; Berthold Hinzen; Stuart J Ince; Steven V Ley



A simple and efficient approach to 1,3-polyols: application to the synthesis of cryptocarya diacetate.  


A highly enantio- and stereoselective synthetic strategy for both syn- and anti-1,3-polyols has been developed. The sequence involves iterative Jacobsen's hydrolytic kinetic resolution (HKR), diastereoselective iodine-induced electrophilic cyclization, and ring-closing metathesis (RCM). This protocol has subsequently been utilized for the synthesis of cryptocarya diacetate, a natural product with broad range of biological activity. PMID:16308881

Kumar, Pradeep; Gupta, Priti; Naidu, S Vasudeva



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

Technology Transfer Automated Retrieval System (TEKTRAN)

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


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

PubMed Central

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

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



Skeletal examination by alizarin staining.  


A skeletal examination of fetuses is required in regulatory embryo-fetal development studies. This chapter describes a method of skeletal examination using alizarin staining. All fetuses are removed from the mother by caesarean section before birth. The fetuses are first examined externally. For larger species (rabbit and minipig), an internal examination can be performed on the fresh soft tissues by microdissection. For smaller species, such as the rat and mouse, half of each litter is fixed for internal soft tissue examination. The other half is used for skeletal examination. A rapid examination of the fresh soft tissues is performed before evisceration and fixation. The staining process takes several days. Following staining, all bones are examined from the head to the tail, in ventral and dorsal positions. PMID:23138906

Reynaud, Lucie; Jocteur-Monrozier, Audrey



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


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

Win, Peter H; Young, Tara A


Development of a Fluorescein Operative Microscope for use During Malignant Glioma Surgery  

Microsoft Academic Search

BackgroundFluorescein has been used in the field of neurosurgery; however, fluorescein enhancement or contrast proved to be inadequate because of a lack of appropriate light sources or filters. A new operative microscope system, in which the microscope itself is equipped with excitation and barrier filters, and the application of this system to surgery for malignant glioma are reported.MethodsBP 450-490, a

Toshihiko Kuroiwa; Yoshinaga Kajimoto; Tomio Ohta



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

PubMed Central

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

Sinnott, Loraine T.



Templating Water Stains for Nanolithography  

PubMed Central

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

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



Stain-Free total protein staining is a superior loading control to ?-actin for Western blots.  


Semi-quantification of proteins using Western blots typically involves normalization against housekeeping genes such as ?-actin. More recently, Ponceau S and Coomassie blue staining have both been shown to be suitable alternatives to housekeeping genes as loading controls. Stain-Free total protein staining offers the advantage of no staining or destaining steps. Evaluation of the use of Stain-Free staining as an alternative to ?-actin or the protein stain Ponceau S showed that Stain-Free staining was superior to ?-actin and as good as or better than Ponceau S staining as a loading control for Western blots. PMID:23747530

Gilda, Jennifer E; Gomes, Aldrin V



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

PubMed Central

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

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



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


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



Fluorescein uptake by a monocarboxylic acid transporter in human intestinal Caco-2 cells.  


The fluorescein transport characteristics of the human intestinal epithelial Caco-2 cell line were examined in monolayer cultures. The initial uptake rate was concentration-dependent and saturable; the Michealis constant and the maximum velocity were 0.40 mM and 1.32 nmol/min/mg protein, respectively. A protonophore, carbonyl cyanide m-chlorophenyl-hydrazone, reduced uptake significantly. The replacement of extracellular sodium ions by lithium ions did not alter the initial uptake rate. These facts imply that the transport is driven by a proton gradient. The initial uptake rate was strongly dependent upon extracellular pH, and the uptake was optimal at approximately pH 5.5. Based on the protolytic constants, the main species of fluorescein in the pH range of 5.5 to 6.0 was calculated to be a monoanion, suggesting that fluorescein was taken up by Caco-2 cells as a monocarboxylate. The following findings support this conclusion: the uptake was inhibited significantly by monocarboxylate compounds such as salicylate and pravastatin, but not by di- or tricarboxylic acids or by acidic amino acids. Furthermore, salicylate-preloaded cells showed remarkably enhanced uptake of fluorescein, indicating that monocarboxylates and fluorescein share a common transport carrier. The transporter has a wide spectrum of substrate recognition and seems likely to be different from MCT1. PMID:11754877

Kuwayama, Kenji; Miyauchi, Seiji; Tateoka, Ryoko; Abe, Hiroshi; Kamo, Naoki



Green One-Pot Synthesis of 2H-Pyrans Under Solvent-Free Conditions Catalysed by Ethylenediammonium Diacetate  

Microsoft Academic Search

Ethylenediammonium diacetate readily catalyses the Knoevenagel-type condensation between 1,3-dicarbonyl substrates and ?,?-unsaturated aldehydes, at room temperature under solvent-free conditions. This rapid, efficient and convenient one-pot approach to the synthesis of 2H-pyrans stands as a significant advance over previously reported protocols. This environmentally friendly methodology has been successfully applied to the synthesis of biologically active natural products of interest zanthosimuline, N-methylflindersine

Martín J. Riveira; Mirta P. Mischne



Adsorption and removal of As(V) and As(III) using Zr-loaded lysine diacetic acid chelating resin  

Microsoft Academic Search

An adsorption process for the removal of As(V) and As(III) was evaluated under various conditions using zirconium(IV) loaded chelating resin (Zr-LDA) with lysine-N?,N? diacetic acid functional groups. Arsenate ions strongly adsorbed in the pH range from 2 to 5, while arsenite was adsorbed between pH 7 and 10.5. The sorption mechanism is an additional complexation between arsenate or arsenite and

Tatineni Balaji; T. Yokoyama; Hideyuki Matsunaga



Evaluation of the probes 2?,7?-dichlorofluorescin diacetate, luminol, and lucigenin as indicators of reactive species formation  

Microsoft Academic Search

This study attempts to provide a critical assessment of three different common approaches to identifying reactive species formed in biological systems: the 2?,7?-dichlorofluorescin diacetate (DCFH-DA) assay, and the luminol- and lucigenin-amplified chemiluminescence assays. There have been several contradictory reports about the specificity of these methods. Our results show that DCFH is oxidized to the fluorescent compound 2?,7?-dichlorofluorescin (DCF) in human

Oddvar Myhre; Jannike M. Andersen; Halvor Aarnes; Frode Fonnum



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

SciTech Connect

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

Zhutaeva, G.V.; Makarova, E.V.; Tkhi Khan, F. [A.N. Frukim Institute of Electrochemistry, Moscow (Russian Federation)] [and others



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


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-DA-Fe(III): (Bis [N,N-bis (carboxymethyl) aminomethyl] fluorescein-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-DA-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 ((1)O2) and hydroxyl radicals (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. PMID:23583854

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



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


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

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



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



Double-Staining Epifluorescence Technique to Assess Frequency of Dividing Cells and Bacteriovory in Natural Populations of Heterotrophic Microprotozoa †  

PubMed Central

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

Sherr, Evelyn B.; Sherr, Barry F.



Problem Solving: Pencil Box Staining  

NSDL National Science Digital Library

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

Boston, Wghb



Modelling ocular pharmacokinetics of fluorescein administered as lyophilisate or conventional eye drops  

Microsoft Academic Search

Objective  The objective of this evaluation was to model ocular pharmacokinetics of fluorescein administered as conventional eye drops\\u000a and as lyophilisate to healthy volunteers in order to assess the relative bioavailability of the lyophilisate formulation.\\u000a \\u000a \\u000a \\u000a Methods  A total of 44 healthy subjects received equivalent doses of fluorescein as lyophilisate to one eye and as eye drops to the\\u000a fellow eye in three

Khaled Abduljalil; Michael Diestelhorst; Oxana Doroshyenko; Anja Lux; Andre Steinfeld; Sven Dinslage; Richard Süverkrüp; Uwe Fuhr



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


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

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



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

National Technical Information Service (NTIS)

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

H. C. Hopps R. C. Brown



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



BAM R14: Clark's Flagellar Stain  

Center for Food Safety and Applied Nutrition (CFSAN)

... To determine staining time (after 2-3 days refrigeration at 4°C), stain a known flagellated organism on 3 or more cleaned slides for various times (eg ... More results from


Methods for chromosome-specific staining  


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.



Mass spectral compatibility of four proteomics stains.  


With the recent introduction of new fluorescence stains to the proteomics market, there is now more choice available. SYPRO Ruby, LavaPurple, Flamingo, and Krypton total protein stains were compared for ease of use, image quality, and compatibility with protein identification by peptide mass fingerprinting (PMF) (MALDI-TOF). All four stains produced good images but with slightly different staining patterns. SYPRO was found to inhibit identification of cysteine and tryptophan containing peptides, which reduced protein identification. PMID:17929854

Ball, Malcolm S; Karuso, Peter



Maintenance of Platelet Viability after Platelet-Labeling with Fluorescein Isothiocyanate  

Microsoft Academic Search

Possible effects of an ex vivo fluorescent labeling procedure of blood platelets on the platelet viability were examined by different function tests. The degree of the impairment of the platelet function proved to be principally correlated with the dye concentration used during the labeling procedure. However, the physiological properties of the labeled platelets obviously remain unrestricted by using intraplatelet fluorescein

J. Klaverkamp; K.-P. Völkl



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

NASA Astrophysics Data System (ADS)

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

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



Fluorescein mercuric acetate—A novel sensor for oral malodour detection  

Microsoft Academic Search

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

Sujatha Jayaraman; Ritu Walia; Nethaji Alagirisamy



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

Microsoft Academic Search

We measured absorption and emission spectra, fluorescence quantum yield, anisotropy, fluorescence resonance energy transfer (FRET), and melting temperature to characterize fluorescein- and tetramethylrhodamine (TMR)-labeled oligonucleotides in solution and when hybridized to a common DNA template. Upon hybridization to the template, both the absorption and emission spectra of TMR-labeled duplexes exhibited a shift with respect to those of labeled oligonucleotides, depending

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



Holographic relaxation spectroscopic study on the structure of gelatin gel doped with fluorescein as a tracer  

Microsoft Academic Search

The structures of gelatin gels have been studied by holographic relaxation spectroscopy (HRS) with fluorescein as a doped tracer. An HRS spectrum with double peaks has been observed. It has been experimentally proven that this “anomalous” HRS spectrum is related to the structures of the gelatin gels. It is shown that two kinds of gel networks are formed when a

Chi Wu; W. Schrof; D. Lilge; E. Luddecke; D. Horn



Differential staining of plant chromosomes with Giemsa  

Microsoft Academic Search

Simple Giemsa staining techniques for revealing banding patterns in somatic chromosomes of plants are described. The value of the methods in the recognition of heterochromatin was demonstrated using five monocotyledonous and two dicotyledonous species. In Trillium grandiflorum the stronger Giemsa stained chromosome segments were shown to be identical with the heterochromatic regions (H-segments) revealed by cold treatment. Preferential staining of

Dieter Schweizer



An Accelerated Method for Staining Tularemia Bacteria.  

National Technical Information Service (NTIS)

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

R. I. Kudelina



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

Microsoft Academic Search

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

Igor Klatzo; Jaime Miquel; Richard Otenasek



Resorption of insoluble, heterologous, fluorescein-collagen sponges in sensitized and non-sensitized rats.  

PubMed Central

Sponges of insoluble bovine collagen were slowly resorbed over a 35-day period when implanted under the back skin of rats. The cellular picture was typical of a mild foreign-body reaction. The reaction to fluorescein-labelled collagen sponges was similar but there was evidence also of a weak immunological response. An acute inflammatory reaction with massive oedema was elicited when fluorescein-labelled collagen sponges were implanted in rats previously sensitized to either fluorescein-collagen or fluorescein-bovine serum albumin. The early invasion by PMN leucocytes subsided after 4 days and caused no observable breakdown of the sponge. The implanted material was rapidly encapsulated by fibrous tissue which was then resorbed along with the sponge between the 7th and 12th day. Macrophages were very active in the sponge at this time, sometimes forming giant cells. Fibroblasts were invading from the periphery with the development of the granulation tissue. The small residue which remained after this time was overrun by granulation tissue and was slowly resorbed up to the 35th day. Throughout the period of study there was only a weak local immunological response after the 28th day. The level of circulating antibodies against the fluorescein hapten was high, but the titre for the antibodies against bovine collagen remained low. The significance of these findings in the pathological destruction of connective tissue is discussed. Images Fig. 1 Fig. 2 Fig. 3a Fig. 3b Fig. 3c Fig. 3d Fig. 3e Fig. 4a Fig. 4b

Etherington, D. J.; Silver, I. A.; Restall, D. J.



Environmentally safe parasitology fixative and stain  

US Patent & Trademark Office Database

A fixative-stain system, which gives superior preservation of nuclear detail, is free from toxic mercury compounds, and which is simple and easy to use, includes a zinc salt and a cobalt salt, in combination, as a fixative, and at least one of Chlorazol Black E, Fast Green FCF and May-Grunwald stains, and preferably the three in admixture, as a staining composition. The fixative may also be used alone. The present fixative-stain system is suitable for fixing and staining all types of parasites such as enteric and other parasites which infect animals and humans.



Quantitative studies of immunofluorescent staining  

PubMed Central

The antiperinuclear factor (APF) is found in a high percentage of sera from patients with rheumatoid arthritis. It can be demonstrated by direct immunofluorescence using the keratohyaline granules of human buccal mucosa as antigenic substrate. Mixing of some normal goat sera with an APF positive serum from a patient with rheumatoid arthritis resulted in an inhibition of the APF titre of the patient's serum. However, there was no clear cut correlation between the APF-positivity of normal goat sera and their inhibitory effect on the APF-reactivity of a human rheumatoid arthritis patient's serum. In reciprocal screening tests the human rheumatoid arthritis serum blocked only one of the APF-reactive goat sera. The reciprocal blocking activity of this goat serum and the patient's serum could be more exactly evaluated by the use of chessboard titrations in an indirect immunofluorescence blocking test. This test consisted of mixing equal volumes of serial dilutions of a goat serum and the patient's serum and subsequent examination of the mixtures for APF using an anti-human IgG conjugate and an anti-goat immunoglobulin conjugate, respectively. The results point to an antibody nature for the APF in preimmune, normal goat sera and to the value of chessboard titrations of this type in demonstrating the identity, non-identity, partial identity (or very close proximity of antigenic determinants) of the antibodies in different antisera which cannot be distinguished by their immunofluorescent staining patterns. ImagesFIG. 1FIG. 2

Wick, G.; Beutner, E. H.



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.



Kinetics of fluorescein decay and its application as a geothermal tracer  

SciTech Connect

This paper reports on fluorescent which is a dye used to trace the path of injected fluids through geothermal reservoirs. The authors have measured its thermal stability at temperatures up to 300{degrees} C in hydrothermal autoclaves at various fluid compositions, pHs, and oxygen concentrations. The results of these experiments indicate that fluorescein will decay less than 10% during a one month tracer test in geothermal reservoirs with temperatures below 210{degrees} C. For tracer test involving longer times and/or higher temperatures, the activation parameters presented in this study can be used to correct for thermal decay. These parameters were applied to a tracer test conducted at the Dixie Valley, Nevada geothermal system to correct for the thermal decay of fluorescein and to deduce the effective temperature of the injection-production flow path.

Adams, M.C.; Davis, J. (Univ. of Utah Research Inst., Salt Lake City, UT (US))



Detection of hyaluronidase activity using fluorescein labeled hyaluronic acid and Fluorescence Correlation Spectroscopy.  


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

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



A new protein conformation indicator based on biarsenical fluorescein with an extended benzoic acid moiety.  


We demonstrate herein a new protein conformation indicator based on biarsenical fluorescein with an extended benzoic acid moiety. The present indicator is reactive to a genetically introduced tetracysteine motif (Cys-Cys-Xaa-Xaa-Cys-Cys, where Xaa is a noncysteine amino acid) of proteins. Compared to the original biarsenical fluorescein (FlAsH) and the biarsenical Nile red analogue (BArNile), the present indicator exhibited larger fluorescence intensity changes in response to Ca(2+)-induced conformational rearrangements of calmodulin. A calculation of the highest occupied molecular orbital (HOMO) level of the benzoic acid moiety of the indicator molecule supports possible involvement of a photoinduced electron transfer (PET) process. These results indicate that the present indicator is useful for sensitive detection of protein conformational changes. PMID:15055950

Nakanishi, Jun; Maeda, Mizuo; Umezawa, Yoshio



Antimicrobial activity of lactic acid bacteria against Listeria monocytogenes on frankfurters formulated with and without lactate/diacetate.  


Contamination by Listeria monocytogenes has been a constant public health threat for the ready-to-eat (RTE) meat industry due to the potential for high mortalities from listeriosis. Lactic acid bacteria (LAB) have shown protective action against various pathogenic bacteria. The aim of this study was to evaluate the antilisterial activity of a combination of three LAB strains (Lactiguard®) on L. monocytogenes. The combination of the LAB was inhibitory to L. monocytogenes inoculated onto frankfurters not containing lactate/diacetate after 8weeks of refrigerated storage (0.6 log reduction compared to L. monocytogenes only control), and when a cell free extract (CFS) of the LAB was added with LAB even more inhibition was obtained (1.2 log reduction compared with L. monocytogenes only). In frankfurters containing lactate/diacetate the LAB and the LAB plus CFS were more effective in reducing growth of L. monocytogenes after 8 weeks of refrigerated storage (2 and 3.3 log reductions respectively). PMID:22704134

Koo, Ok-Kyung; Eggleton, Mallory; O'Bryan, Corliss A; Crandall, Philip G; Ricke, Steven C



Stain Removal from a Silicone Maxillofacial Elastomer  

Microsoft Academic Search

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

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



Contextual detection of ischemic regions in ultra-wide-field-of-view retinal fluorescein angiograms  

Microsoft Academic Search

We report a novel prototype algorithm using contextual knowledge to locate ischemic regions in ultra- wide-field-of-view retinal fluorescein angiograms. We use high- resolution images acquired by an Optos ultra-wide-field-of- view (more than 200 degrees) scanning laser ophthalmoscope. We leverage the simultaneous occurrence of ischemia with a number of other signs, detected automatically, typical for the state of progress of the

E. Trucco; C. R. Buchanan; T. Aslam; B. Dhillon



X-ray photoelectron spectroscopy of fluorescein adsorbed on model solar-cell surfaces  

Microsoft Academic Search

We report on the X-ray photoelectron spectroscopy of fluorescein molecules adsorbed under UHV conditions on single crystal rutile TiO2(110) and Au\\/TiO2(110) surfaces. The molecule is thought to bond covalently to these model solar-cell surfaces primarily through the deprotonated carboxylic group, with an additional interaction arising between the triple ring structure of the molecule and the surface.

James N O’Shea; J Ben Taylor; Emily F Smith



Silver staining of proteins in polyacrylamide gels  

PubMed Central

Silver staining is used to detect proteins after electrophoretic separation on polyacrylamide gels. It combines excellent sensitivity (in the low nanogram range) whilst using very simple and cheap equipment and chemicals. It is compatible with downstream processing such as mass spectrometry analysis after protein digestion. The sequential phases of silver staining are protein fixation, then sensitization, then silver impregnation and finally image development. Several variants of silver staining are described here, which can be completed in a time range from 2 hours to one day after the end of the electrophoretic separation. Once completed, the stain is stable for several weeks

Chevallet, Mireille; Luche, Sylvie; Rabilloud, Thierry



Bodian's Silver Method Stains Neurofilament Polypeptides  

NASA Astrophysics Data System (ADS)

Bodian's silver method was used to stain polypeptides of rat spinal cord or peripheral nerve separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The bands corresponding to the three polypeptide subunits of the neurofilaments were intensely impregnated. Two other polypeptides were stained inconsistently and less intensely. The tubulin band was stained weakly or not at all; other polypeptides, including glial fibrillary acidic protein, actin, and vimentin, remained unstained. This novel application of Bodian's method provides indirect proof that neurofilaments are the neuronal subcellular structure stained by the technique.

Gambetti, P.; Autilio-Gambetti, L.; Papasozomenos, S. Ch.



Selective labeling of histidine by a designed fluorescein-based probe.  


The synthesis of a novel fluorescent probe, 3-epoxypropoxy fluorescein (EPF), and its properties for labeling of histidine are described. The probe contained a fluorescein fluorophore with long-wavelength response and an active epoxy labeling group. In alkaline media EPF reacted selectively with histidine, rather than with other amino acids, causing a large increase in fluorescence intensity and thereby allowing a selective detection of histidine. This fluorescence increase resembled that of the fluorescein diaion with the increase of the media basicity, suggesting that the addition reaction of histidine with the epoxy group provides the fluorophore moiety with a basic molecular environment. As an application of this probe, fluorescent labeling of histidine in human serum was attempted and the obtained results were in agreement with those given by using histidine-nickel complex adsorptive voltammetry. Further, the relative S.D. of the method was 1.8% for 10 replicate determinations of 0.55 muM histidine. When 10muM of EPF was used, the linear range for histidine was 0.007-10muM with a detection limit (S/N=3) of 0.001muM. PMID:18969304

Li, Xiaohua; Ma, Huimin; Dong, Suying; Duan, Xuejun; Liang, Shuchuan



Centripetal movement of fluorescein dextrans in the cornea: relevance to arcus.  


The centripetal movement of fluorescein and fluorescein-labelled dextrans (4 to 150 kD) from sclera or cut edge of the cornea was determined in isolated rabbit corneas at 4 and 24 h. Corneas were divided into 5.5 mm diameter central core, inner 5.5 to 8 mm donut, 8 to 12 mm peripheral donut and, where applicable, scleral rim. For all molecules greater than sodium fluorescein (376 D) tracer concentrations in the 5.5 mm core and the 5.5 to 8 mm donut were equal. Without sclera rim, the more central portions of the cornea (5.5 mm core and 5.5 to 8 mm donut) had tracer concentrations equal to those of corneas-with-sclera for all tracers greater than 10 kD. The tracer concentrations in the central cornea were the same in the presence or absence of sclera. The data indicate a physiological barrier to the lateral diffusion of molecules greater than 10 kD between the peripheral and more central cornea. PMID:2447743

Green, K; DeBarge, L R; Cheeks, L; Phillips, C I



Rational engineering of a fluorescein-binding anticalin for improved ligand affinity.  


The anticalin FluA is an artificial lipocalin with novelspecificity for the fluorescein group, which was engineered from an insect bilin-binding protein by targeted random mutagenesis and selection. Based on the crystal structure of FluA, an attempt was made to improve the complementarity of its ligand pocket to fluorescein by rational protein design. Several side chains participating in sub-optimal interactions with the ligand were identified and replaced by residues that promised a better steric fit. As a result, the substitution of Ala45 by Ile and of Ser114 by Thr or Arg led to a tight affinity of ca. 1 nM, which is approximately 30-fold better than that of the parental anticalin. Similar to the original FluA, the improved version shows almost complete quenching of the bound ligand fluorescence. Interestingly, the quenching effect was significantly reduced when Trp129 was replaced by Tyr, thus supporting the previously postulated role of this residue, which closely packs against the bound ligand, for efficient electron transfer to the excited fluorescein. Circular dichroism spectra revealed that all variants investigated had retained the lipocalin fold. Corresponding thermal unfolding experiments confirmed similar folding stabilities, with melting temperatures ranging from 52.9 to 60.5 degrees C (i.e., for the high-affinity variant). PMID:16307475

Vopel, Sven; Mühlbach, Hermine; Skerra, Arne



Synthesis and structure of the product of bis-condensation of 4-methyl-2,6-diformylphenol with dimethyl N?,N?-hydrazine diacetate  

Microsoft Academic Search

Bis-condensation of 4-methyl-2,6-diformylphenol with dimethyl N?,N?-hydrazine diacetate gave product I. The structure of I\\u000a was determined by X-ray analysis. The conformations of the two side chains of dimethyl N?,N?-hydrazine diacetate are significantly\\u000a different. The position of one chain is fixed by the intramolecular H-bond of O?H.…N type. Due to its conformation, compound\\u000a I is partly ready for complex formation with

V. Kh. Kravtsov; V. I. Lozan; Yu. A. Simonov; O. A. Bologa; N. V. Gerbeleu; T. I. Malinovskii



Purified azure B as a reticulocyte stain  

Microsoft Academic Search

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

P N Marshall; S A Bentley; S M Lewis



Stain removal from a silicone maxillofacial elastomer.  


In this study, environmental stains were removed from maxillofacial elastomers by solvent extraction. Silastic 44210, an RTV silicone with proven color and physical property stability, was stained with lipstick, disclosing solution, and methylene blue. These stains were then removed by solvent extraction with each of four chemically dissimilar solvents, namely: toluene, benzene, 1,1,1-trichloroethane, and n-hexane. An additional series of samples was prepared with 11 maxillofacial pigments, not for staining, but for evaluation of pigment stability. Results obtained from spectrophotometric measurements before and after solvent extraction demonstrated the effectiveness of solvent extraction in removing stains, while there was little or no change in the color of the pigments or the base elastomer. PMID:6944340

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



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

PubMed Central

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

De Carlo, Sacha; Harris, J. Robin



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

Microsoft Academic Search

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

S. A. Nathoo; A. Gaffar



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

Microsoft Academic Search

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

William H. Massover; Philip Marsh



Combined in situ zymography, immunofluorescence, and staining of iron oxide particles in paraffin-embedded, zinc-fixed tissue sections.  


Superparamagnetic iron oxide particles are used as potent contrast agents in magnetic resonance imaging. In histology, these particles are frequently visualized by Prussian blue iron staining of aldehyde-fixed, paraffin-embedded tissues. Recently, zinc salt-based fixative was shown to preserve enzyme activity in paraffin-embedded tissues. In this study, we demonstrate that zinc fixation allows combining in situ zymography with fluorescence immunohistochemistry (IHC) and iron staining for advanced biologic investigation of iron oxide particle accumulation. Very small iron oxide particles, developed for magnetic resonance angiography, were applied intravenously to BALB/c nude mice. After 3 hours, spleens were explanted and subjected to zinc fixation and paraffin embedding. Cut tissue sections were further processed to in situ zymography, IHC, and Prussian blue staining procedures. The combination of in situ zymography as well as IHC with subsequent Prussian blue iron staining on zinc-fixed paraffin-embedded tissues resulted in excellent histologic images of enzyme activity, protease distribution, and iron oxide particle accumulation. The combination of all three stains on a single section allowed direct comparison with only moderate degradation of fluorescein isothiocyanate-labeled substrate. This protocol is useful for investigating the biologic environment of accumulating iron oxide particles, with excellent preservation of morphology. PMID:22954182

Schellenberger, Eyk; Haeckel, Akvile; Schoenzart, Lena; Appler, Franziska; Schnorr, Joerg; Taupitz, Matthias; Hamm, Bernd


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

Microsoft Academic Search

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

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



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

Microsoft Academic Search

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

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



Evidence for substantial maintenance of membrane integrity and cell viability in normally developing grape (Vitis vinifera L.) berries throughout development  

Microsoft Academic Search

Fluorescein diacetate (FDA) was used as a vital stain to assay membrane integrity (cell viability) in meso- carp tissue of the developing grape (Vitis vinifera L.) berry in order to test the hypothesis that there is a substantial loss of compartmentation in these cells during ripening. This technique was also used to determine whether loss of viability was associated with

Mark Krasnow; Mark Matthews; Ken Shackel



Improved development by Taxol pretreatment after vitrification of in vitro matured porcine oocytes  

Microsoft Academic Search

This study was designed to examine the effect of Taxol pretreatment on vitrification of porcine oocytes matured in vitro by an open pulled straw (OPS) method. In the first experiment, the effect of Taxol pretreatment and fluorescein diacetate (FDA) staining on parthenogenetic development of oocytes was evaluated. In the second experiment, viability, microtubule organiz- ation and embryo development of oocytes

Wen-Qing Shi; Shi-En Zhu; Dong Zhang; Wei-Hua Wang; Guo-Liang Tang; Yun-Peng Hou; Shu-Jun Tian



Novel Characteristics of Glutamate-Induced Cell Death in Primary Septohippocampal Cultures: Relationship to Calpain and Caspase3 Protease Activation  

Microsoft Academic Search

Studies examined the phenotypic characteristics of glutamate-induced cell death and their relationship to calpain and caspase-3 activation. Cell viability was assessed by fluorescein diacetate and propidium iodide staining and lactate dehydrogenase release. Calpain and caspase-3 activity was inferred from signature proteolytic fragmentation of ?-spectrin. Characterization of cell death phenotypes was assessed by Hoechst 33258 and DNA fragmentation assays. Exposure of

Xiurong Zhao; Jennifer K. Newcomb; Brian R. Pike; Kevin K. W. Wang; Domenico d'Avella; Ronald L. Hayes



Property of thimerosal-induced decrease in cellular content of glutathione in rat thymocytes: a flow cytometric study with 5-chloromethylfluorescein diacetate  

Microsoft Academic Search

There is a concern on the part of public health community that adverse health consequences by thimerosal, a preservative in vaccines for infants, may occur among infants during immunization schedule. Therefore, the effect of thimerosal on cellular content of glutathione was examined on thymocytes obtained from 4-week-old rats using a flow cytometer and 5-chloromethylfluorescein diacetate. Thimerosal at concentrations ranging from

T Ueha-Ishibashi; T Tatsuishi; K Iwase; H Nakao; C Umebayashi; Y Nishizaki; Y Nishimura; Y Oyama; S Hirama; Y Okano



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...


Preparation and characterization of imino diacetic acid functionalized alginate beads for removal of contaminants from waste water: I. methylene blue cationic dye model  

Microsoft Academic Search

This study deals with the development of a clean and safe process for water pollution remediation. We studied the potential use of Imino Diacetic Acid (IDA) activated calcium alginate beads for removal of cationic dyes from colored effluents in dynamic batch mode. Methylene blue (MB) has been chosen as a dye model for the study. The parameters that affect the

Mohamed Samir Mohy Eldin; Emad Ali Soliman; Ahmed Abdel Fattah Elzatahry; Mohamed Ramadan Elaassar; Marwa Farouk Elkady; Aref Mohamed Abdel Rahman; Mohamed Elsayed Yossef; Bassant Yossri Eweida




Microsoft Academic Search

Phase change materials (PCMs) of cellulose diacetate (CDA) modified with polyethylene glycol (PEG) prepared by chemical bonding and physical blending methods exhibit different characteristics: Chemically bonded materials exhibit typical solid-solid phase change characteristics, but the blended materials exhibit solid-liquid phase change characteristics. In all these materials, CDA is the framework, and PEG is a working substance that gives and takes

En-Yong Ding; Yong Jiang; Guo-Kang Li



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...



Technology Transfer Automated Retrieval System (TEKTRAN)

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



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...



Technology Transfer Automated Retrieval System (TEKTRAN)

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


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


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

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



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

Microsoft Academic Search

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

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



Cochlear microdialysis for quantification of dexamethasone and fluorescein entry into scala tympani during round window administration.  


Before new drugs for the treatment of inner ear disorders can be studied in controlled clinical trials, it is important that their pharmacokinetics be established in inner ear fluids. Microdialysis allows drug levels to be measured in perilymph without the volume disturbances and potential cerebrospinal fluid contamination associated with fluid sampling. The aims of this study were to show: (i) that despite low recovery rates from miniature dialysis probes, significant amounts of drug are removed from small fluid compartments, (ii) that dialysis sampling artifacts can be accounted for using computer simulations and (iii) that microdialysis allows quantification of the entry rates through the round window membrane (RWM) into scala tympani (ST). Initial experiments used microdialysis probes in small compartments in vitro containing sodium fluorescein. Stable concentrations were observed in large compartments (1000 microl) but significant concentration declines were observed in smaller compartments (100, 10 and 5.6 microl) comparable to the size of the inner ear. Computer simulations of these experiments closely approximated the experimental data. In in vivo experiments, sodium fluorescein 10 mg/ml and dexamethasone-dihydrogen-phosphate disodium salt 8 mg/ml were simultaneously applied to the RWM of guinea pigs. Perilymph concentration in the basal turn of ST was monitored using microdialysis. The fluorescein concentration reached after 200 min application (585+/-527 microg/ml) was approximately twice that of dexamethasone phosphate (291+/-369 microg/ml). Substantial variation in concentrations was found between animals by approximately a factor of 34 for fluorescein and at least 41 for dexamethasone phosphate. This is, to a large extent, thought to be the result of the RWM permeability varying in different animals. It was not caused by substance analysis variations, because two different analytic methods were used and the concentration ratio between the two substances remained nearly constant across the experiments and because differences were apparent for the repeated samples obtained in each animal. Interpretation of the results using computer simulations allowed RWM permeability to be quantified. It also demonstrated, however, that cochlear clearance values could not be reliably obtained with microdialysis because of the significant contribution of dialysis to clearance. The observed interanimal variation, e.g., in RWM permeability, is likely to be clinically relevant to the local application of drugs in patients. PMID:16442251

Hahn, Hartmut; Kammerer, Bernd; DiMauro, Andre; Salt, Alec N; Plontke, Stefan K



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

PubMed Central

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

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



Cholinergic Staining of Bronchus Associated Lymphoid Tissue  

Microsoft Academic Search

The cholinergic staining of human bronchus-associated lymphoid tissue (BALT) was studied in humans. Morsels of the human lung (containing BALT) were harvested, after having obtained the appropriate approvals, during autopsies in 24 human subjects. The samples were stained by means of the enzymatic technique of acetylcholinesterase (AChE) and\\/or the monoclonal immunohistochemical method of choline acetyltransferase (ChAT). A morphometrical analysis was

Carlo Cavallotti; Gianfranco Tonnarini; Vito D’Andrea; Daniela Cavallotti



Differential staining of ocular goblet cells  

Microsoft Academic Search

Millipore filters were used to obtain sheets of cells from the ocular surface. Using Periodic Acid Schiff-haematoxylin the intracellular neutral mucus of the goblet cells stains a brilliant, bright pink and the cell nuclei dark blue making it possible to observe the epithelial cells and the goblet cell population.In certain ocular surface diseases the size of the PAS-haematoxylin staining goblet

G G W Adams; P N Dilly; GGW Adams



A 'Magnetic' Gram Stain for Bacterial Detection  

PubMed Central

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

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



Intraoperative tissue staining of invaded oral carcinoma.  


The purpose of this study was to assess the ability of intraoperative tissue staining with consecutive application of 0.4% indigo carmine and 0.5% Congo red to demonstrate the extent and border of oral carcinoma invasion. Seventeen patients were included in the study. Once the oral tumor was resected, a vertical section of surgical specimen was taken from the central part of the tumor. The extent and border of the invaded carcinoma were assessed on digital microscopic examination with tissue staining. The results of assessments were compared with corresponding results of conventional histopathological analysis with HE staining, which is considered the gold standard. Tissue staining produced a brown-black stain on normal muscle, connective, and salivary tissues but not tumor and epithelial tissues. It clearly demonstrated the extent and border of tumor invasion in 13 of 17 patients (76.5%); however, detection of remnant vital tumor cells in scar tissue after neoadjuvant chemotherapy, and distinction between the tumor and adipose tissue scattered in the muscle tissue was difficult. The results of this study showed that intraoperative tissue staining was a possible method in demonstrating the extent and border of carcinoma deeply invaded in the soft tissue and selecting the site for additional frozen section analysis, although the method needed some refinement. PMID:18575826

Kurita, Hiroshi; Kamata, Takahiro; Koike, Takeshi; Kobayashi, Hiroichi; Kurashina, Kenji



RERBEE: robust efficient registration via bifurcations and elongated elements applied to retinal fluorescein angiogram sequences.  


We present RERBEE (robust efficient registration via bifurcations and elongated elements), a novel feature-based registration algorithm able to correct local deformations in high-resolution ultra-wide field-of-view (UWFV) fluorescein angiogram (FA) sequences of the retina. The algorithm is able to cope with peripheral blurring, severe occlusions, presence of retinal pathologies and the change of image content due to the perfusion of the fluorescein dye in time. We have used the computational power of a graphics processor to increase the performance of the most computationally expensive parts of the algorithm by a factor of over × 1300, enabling the algorithm to register a pair of 3900 × 3072 UWFV FA images in 5-10 min instead of the 5-7 h required using only the CPU. We demonstrate accurate results on real data with 267 image pairs from a total of 277 (96.4%) graded as correctly registered by a clinician and 10 (3.6%) graded as correctly registered with minor errors but usable for clinical purposes. Quantitative comparison with state-of-the-art intensity-based and feature-based registration methods using synthetic data is also reported. We also show some potential usage of a correctly aligned sequence for vein/artery discrimination and automatic lesion detection. PMID:21908251

Perez-Rovira, Adria; Cabido, Raul; Trucco, Emanuele; McKenna, Stephen J; Hubschman, Jean Pierre



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


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



Staining efficiency of specific proteins depends on the staining method: wheat gluten proteins.  


To analyze gluten proteins involved in celiac disease (CD) by proteomic analysis, prolamins extracted from hexaploid wheat varieties were analyzed by SDS-PAGE and 2-DE. Differences between staining methods (CBB, silver nitrate, SYPRO Ruby, and CyDye) were analyzed in comparison to immunoblotting. Staining efficiency varied per protein across methods, and complete staining of all gluten proteins could not be achieved by one of these methods. Care should be taken in the selection of staining method especially if one wants to relate the results to data obtained by immunoblotting. PMID:18398878

van den Broeck, Hetty C; America, Antoine H P; Smulders, Marinus J M; Gilissen, Ludovicus J W J; van der Meer, Ingrid M



Stain Removal from a Pigmented Silicone Maxillofacial Elastomer  

Microsoft Academic Search

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

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



Rapid coomassie blue staining of protein gels.  


Coomassie brilliant blue R250 (CBR-250) and silver staining are the most widely used methods for the routine visualization of proteins separated by SDS-PAGE. CBR-250 is an organic dye that complexes with basic amino acids, such as arginine, lysine, and histidine, as well as tyrosine. Conventional CBR-250 staining is capable of detecting as little as 30-100 ng of protein, but sensitivity can be improved by performing the staining and destaining at elevated temperatures. The method described in this protocol, which is a modified version of the conventional Coomassie protocol, speeds up the destaining process for faster results with increased sensitivity and is compatible with mass-spectrometry-based methods for identifying proteins. PMID:20360367

Simpson, Richard J



Mast cell ultrastructure and staining in tissue.  


Mast cells are bone marrow-derived cells that are widely distributed in the tissue. They are found predominantly in the subepithelial tissue near blood vessels and nerves and usually are sprinkled diffusely without forming clusters. In tissue sections stained with hematoxylin and eosin, normal mast cells usually display a round-to-oval nucleus with clumped chromatin and indistinct or no nucleoli. They have moderately abundant cytoplasm and are oval, spindle, or polygonal in shape. The cytoplasm is amphophilic, and sometimes small slightly eosinophilic granules may be visible. Hematoxylin and eosin staining is not a specific or reliable method for detecting mast cells in tissue sections because of variable cellular morphology. For confirmation of mast cells, special stains, such as mast cell tryptase or CD117, are required. PMID:16110149

Shukla, Shruti A; Veerappan, Ranjitha; Whittimore, Judy S; Ellen Miller, Lou; Youngberg, George A



CD31 staining in epithelioid sarcoma.  


We report an unusual case of epithelioid sarcoma. The tumour occurred in the finger of a 27-year-old female. The clinical history, histology and the electron microscopy of the lesion were typical for epithelioid sarcoma. However, immunohistochemical analysis showed strong membranous CD31 staining, a finding hitherto not described. All other robust vascular markers, including factor-VIII-related antigen (FVIIIrag) were negative. The findings were compared with the available literature data, leading us to conclude that there is insufficient evidence for endothelial derivation of epithelioid sarcoma, but in the differential diagnosis with vascular tumours CD31 may stain and to rule out angiosarcoma FVIIIrag is a useful antibody. PMID:12743818

den Bakker, M A; Flood, S J; Kliffen, M



Detection Of Concrete Deterioration By Staining  


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)



21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2010 CFR

...2009-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food...HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains. (a) Identification. Dye and...



21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2010 CFR

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



Resistance to Extrinsic Stains by Hydrophobic Composite Resin Systems  

Microsoft Academic Search

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

W. H. Douglas; R. G. Craig



Pleural and Pulmonary Staining at Inferior Phrenic Arteriography Mimicking a Tumor Staining of Hepatocellular Carcinoma  

SciTech Connect

Purpose: To describe the findings of pleural and pulmonary staining of the inferior phrenic artery, which can be confused with tumor staining during transarterial chemoembolization (TACE) of hepatoma.Methods: Fifteen patients who showed pleural and pulmonary staining without relationship to hepatic masses at inferior phrenic arteriography were enrolled. The staining was noted at initial TACE (n = 8), at successive TACE (n = 5), and after hepatic surgery (n = 2). The angiographic pattern, the presence of pleural change on computed tomography (CT), and clinical history were evaluated.Results: Draining pulmonary veins were seen in all cases. The lower margin of the staining corresponded to the lower margin of the pleura in 10 patients. CT showed pleural and/or pulmonary abnormalities in all cases. After embolization of the inferior phrenic artery, the accumulation of iodized oil in the lung was noted.Conclusion: Understanding the CT and angiographic findings of pleural and pulmonary staining during TACE may help differentiate benign staining from tumor staining.

Lee, Deok Hee; Hwang, Jae Cheol; Lim, Soo Mee; Yoon, Hyun-Ki; Sung, Kyu-Bo [Department of Diagnostic Radiology, Asan Medical Center, University of Ulsan College of Medicine, 388-1 Poong-nap Dong, Song-pa Gu, Seoul 138-040 (Korea, Republic of); Song, Ho-Young [Department of Diagnostic Radiology, Kangnung Hospital, 415 Bang-dong Ri, Sa-cheon Myeon, Kang-nung Si, Kang-won Do 210-711 (Korea, Republic of)



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

SciTech Connect

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

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



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.



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

PubMed Central

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

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



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


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

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



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


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



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))



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


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



Effect of Ovulen-50(ethynodiol diacetate 1 mg and ethynyloestradiol 0,05 mg) on protein, nucleic acids and nucleases in the rat liver.  


Treatment of adult female rats with Ovulen-50 (ethynodiol diacetate 1.0 mg and ethynyloestradiol 0.05 mg--one tenth tablet per day) led to a small but significant fall in total protein, RNA and DNA content of the liver. Alkaline and acid DNAse activity also tended to decline, whereas alkaline RNAase tended to show a marginal rise after twentyfour days of treatment. PMID:1010688

Mukundan, M A; Bamji, M S



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



A magnetic Gram stain for bacterial detection.  


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

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



Transformed steroids. Communication 154. Production of a 17-spirooxetan-20-one under conditions of 21-hydroxylation of 16. cap alpha. -acetamidopregn-5-ene-3. beta. ,17. cap alpha. -diol-20-one with iodosobenzene diacetate  

SciTech Connect

The possibility of the one-step synthesis of 17-spirooxetan-20-ones from 17..beta..-acetyl-17..cap alpha..-hydroxy steroids with the aid of iodosobenzene diacetate in a methanol solution of alkali was demonstrated.

Kamernitskii, A.V.; Fadeeva, T.M.; Turuta, A.M.



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


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

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



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.



Localization of viral antigen in narcissus leaves infected with yellow stripe virus, determined by means of a fluorescein conjugated antiserum  

Microsoft Academic Search

By means of a fluorescein conjugated antiserum, local aggregation of antigenic material in narcissus leaves infected with yellow stripe virus was demonstrated in the cytoplasm of epidermal and parenchyma cells. By comparing results obtained by fluorescent-microscopy with those obtained by light and electron microscopy it is reasonable to assume that the antigenic material consists of aggregates of virus particles.

Margaretha C. Cremer; J. A. Van Der Veken



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.



Staining of water trees with methylene blue explained  

Microsoft Academic Search

Staining with a methylene blue solution, a popular technique for achieving the required high contrast images of water trees, was carried out at 70°C. It is shown that methylene blue stains the complete tree, which justifies water tree length measurements after staining. The water tree is not stained exclusively, but rather faster and probably to a higher degree than the

R. Ross; J. J. Smit; P. Aukema



Meconium staining of the brainstem with open myelomeningocele.  


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

Lam, Sandi; Grandhi, Ramesh; Greene, Stephanie



Staining and evaluation of serum lipoproteins  

Microsoft Academic Search

Summary  A simple and reliable method of prestaining lipoproteins using a solution of Sudan blue in propane 1:2 diol prior to paper\\u000a electrophoresis is described. Simultaneous examination of sera by prestaining and staining after electrophoresis indicate\\u000a that prestaining is a reliable method for investigating alpha and beta lipoproteins, but not for determining the neutral fat\\u000a present in the serum.\\u000a \\u000a A comparison

Robert G. McFarlane



Activity staining of endoglucanases in polyacrylamide gels.  


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

Mathew, R; Rao, K K



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


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


The effects and interactions of temperature (56.3-60 °C), sodium lactate (SL; 0-4.8%), sodium diacetate (SDA; 0-2.5%), and pediocin (0-10,000 AU) on starved Listeria monocytogenes (10(7) CFU/g) on bologna were investigated. Bologna slices containing SL and SDA in the formulation were dipped in pediocin, surface inoculated, and treated at various temperatures using combinations of parameters determined by central composite design. D-values were calculated. The observed D-values ranged from 2.8 min at 60 °C to 24.61 min at 56.3 °C. Injury ranged from 9.1 to 76% under various conditions. The observed D-values were analyzed using second order response surface regression for temperature, SL, SDA, and pediocin, and a predictive model was developed. Predicted D-values were calculated and ranged from 3.7 to 19 min for various combinations of parameters. Temperature alone reduced the predicted D-values from 33.96 min at 56.3 °C to 11.51 min at 60 °C. Addition of SL showed a protective effect. Other combination treatments either reduced or increased D-values depending on temperature. The combination of SL and SDA was effective at lower temperatures, however, higher levels of SDA at higher temperatures made the organism more heat resistant. Pediocin (up to 5000 AU) with increasing temperature and SDA reduced D-values. Depending on temperature and concentration, the interactions between various additives can affect thermal inactivation of L. monocytogenes on bologna. Starvation rendered L. monocytogenes more susceptible to heat and additives. PMID:21356449

Grosulescu, Camelia; Juneja, Vijay K; Ravishankar, Sadhana



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


The effects and interactions of temperature (56.3-60 degrees C), sodium lactate (SL; 0-4.8%), sodium diacetate (SD; 0-0.25%) and pediocin (0-10,000 AU) on Listeria monocytogenes on bologna were studied and a predictive inactivation model was developed. Bologna was manufactured with different SL/SD concentrations in the formulation, dipped in pediocin solution and treated at different temperatures using combinations of parameters determined by central composite design. D-values were calculated and analyzed using second order response regression. Predicted D-values were also calculated. The observed D-values for L. monocytogenes on bologna ranged from 2.10 to 35.59 min. Temperature alone decreased predicted D-values from 99.02 min at 56.3 degrees C to 44.71 min at 60.0 degrees C. Adding SL decreased D-values (85.43-22.71 min) further; however, heat and SD combined was the most effective for reducing L. monocytogenes on bologna. An SD level of 0.25% at 58.2 degrees C had the overall lowest predicted D-value (15.95 min). Combination treatments increased or decreased D-values, depending on the temperature. Pediocin (2500 and 5000 AU) and heat decreased D-values, but exhibited a protective effect at higher concentrations (>or=7500 AU). The results showed that interactions between additives in formulations can vary at different temperatures/concentrations, thereby affecting thermal inactivation of foodborne pathogens in meat products. Hence, food processors should modify food formulations carefully, and verify with adequate testing so that product safety is not compromised. PMID:19913694

Maks, Nicole; Zhu, Libin; Juneja, Vijay K; Ravishankar, Sadhana



Effects of ortho-phthalaldehyde, glutaraldehyde and chlorhexidine diacetate on Mycobacterium chelonae and Mycobacterium abscessus strains with modified permeability.  


The mechanisms of the mycobactericidal action of ortho-phthalaldehyde (OPA), glutaraldehyde (GTA) and chlorhexidine diacetate (CHA) were investigated using mycobacterial spheroplasts of two reference strains, Mycobacterium chelonae NCTC 946, Mycobacterium abscessus NCTC 10882 and two GTA-resistant strains, M. chelonae Epping and M. chelonae Harefield. Transmission electron microscopy of the spheroplasts revealed an altered cell wall structure compared with the parent cells. Structural alterations resulting from the spheroplasting process were in part correlated to a loss of lipid content. Low concentrations of CHA induced protein coagulation in M. chelonae NCTC 946 spheroplasts, which also exhibited the highest loss of free non-polar lipids. Higher concentrations of CHA were required to produce similar results to the other spheroplasts investigated in which there was a less substantial decrease in lipid content. OPA (0.5% w/v) readily penetrated the residual cell wall and cytoplasmic membrane, producing significant protein coagulation in M. chelonae NCTC 946. GTA (0.5% v/v) induced a similar effect but to a lesser extent. Pre-treatment of the spheroplasts with OPA and GTA and their subsequent suspension in water demonstrated that GTA was a more potent cross-linking agent. This protective effect of GTA results from extensive cross-linking of amino and/or sulphydryl side-chain groups of proteins. The rapid mycobactericidal effect of OPA probably arises from its more efficient penetration across biological membranes. Mycobacterial spheroplasts represented a useful cellular model with an altered cell wall permeability. This study also showed the importance of the mycobacterial cell wall in conferring intrinsic resistance to CHA. PMID:12615857

Fraud, S; Hann, A C; Maillard, J-Y; Russell, A D



Radiation (gamma) resistance and postirradiation growth of Listeria monocytogenes suspended in beef bologna containing sodium diacetate and potassium lactate.  


Listeria monocytogenes, a psychrotrophic foodborne pathogen, is a frequent postprocessing contaminant of ready-to-eat (RTE) meat products, including frankfurters and bologna. Ionizing radiation can eliminate L. monocytogenes from RTE meats. When they are incorporated into fine-emulsion sausages, sodium diacetate (SDA) and potassium lactate (PL) mixtures inhibit the growth of L. monocytogenes. The radiation resistance of L. monocytogenes, and its ability to proliferate during long-term refrigerated storage (9 degrees C), when inoculated into beef bologna that contained 0% SDA-0% PL, 0.07% SDA-1% PL, and 0.15% SDA-2% PL, were determined. The radiation doses required to eliminate 90% of the viable L. monocytogenes cells were 0.56 kGy for bologna containing 0% SDA-0% PL, 0.53 kGy for bologna containing 0.07% SDA-1% PL, and 0.46 kGy for bologna containing 0.15% SDA-2% PL. L. monocytogenes was able to proliferate on bologna containing 0% SDA-0% PL during refrigerated storage, but the onset of proliferation was delayed by the addition of the SDA-PL mixtures. An ionizing radiation dose of 3.0 kGy prevented the proliferation of L. monocytogenes and background microflora in bologna containing 0.07% SDA-1% PL and in bologna containing 0.15% SDA-2% PL over 8 weeks of storage at 9 degrees C. Little effect on lipid oxidation and color of the control bologna, or bologna containing SDA-PL mixtures, was observed upon irradiation at either 1.5 or 3.0 kGy. PMID:14627282

Sommers, Christopher; Fan, Xuetong; Niemira, Brendan A; Sokorai, Kimberly



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.



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

Microsoft Academic Search

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

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



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.

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



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.

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



Synthesis, biological evaluation, and in vivo imaging of the first camptothecin-fluorescein conjugate.  


The first synthesis and photophysical properties of a fluorecently labeled camptothecin derivative, namely, camptothecin-FI (CPT-FI), an antitumoral agent that targets topoisomerase I, are reported. The preparation of this fluorescent conjugate is based on a highly convergent and flexible approach which enables the rapid chemical modification of the AB ring system of this fragile pentacyclic alkaloid, aimed at introducing an anchoring point to graft the fluorophore. The selection of a fluorescein analogue as the reporter group has enabled us to get the first green-emitting CPT conjugate exhibiting valuable spectral properties and retaining biological properties of native CPT. Indeed, in biological models, i.e., glioma cell lines U87 and/or T98, the kinetics of cell endocytosis, as well as the efficacy of CPT-FI were compared to those of CPT. CPT-FI fluorescence was measured in the cytosolic compartment of T98 glioma cells from 5 min treatment and remained detectable until 48 h. As CPT, CPT-FI drastically inhibited glioma growth and cell cycle but exhibited a reduced affinity as compared to the native CPT. In vivo and ex vivo imaging studies of CPT-FI intratumoraly injected into a model of NIH-3T3 murine tumor xenografts in nude mice, showed accumulation around the injected site area, which is very promising to target tumors and follow biodistribution in vivo. PMID:23750546

Chevalier, Arnaud; Dubois, Martine; Le Joncour, Vadim; Dautrey, Sébastien; Lecointre, Céline; Romieu, Anthony; Renard, Pierre-Yves; Castel, Hélène; Sabot, Cyrille



Saturable CD14-dependent binding of fluorescein-labeled lipopolysaccharide to human monocytes.  

PubMed Central

We used rough lipopolysaccharide (ReLPS) to construct a fluorescein-labeled LPS (FITC-LPS) with a very high labeling efficiency that bound to isolated human monocytes in a CD14-dependent fashion and that in this respect behaved indistinctively from native LPS. The CD14-dependent binding could be inhibited either by a 1,000-fold excess of unlabeled LPS or by polymyxin B, bactericidal/permeability-increasing protein, cationic protein 18, or soluble CD14. Although this FITC-LPS preparation no longer possessed the ability to prime neutrophils for the production of reactive oxygen species or to stimulate human monocytes to produce tumor necrosis factor, activation of the Limulus amoebocyte lysate cascade was comparable to activation by native LPS. Binding to monocytes was enhanced by human pooled serum (HPS) or LPS-binding protein (LBP) for LPS concentrations up to 100 ng/ml and was completely CD14 dependent. For LPS concentrations exceeding 100 ng/ml, binding was still partially CD14 dependent, but not HPS or LBP dependent. CD14-dependent association of LPS with monocytes was shown to be totally saturable. In conclusion, we found an HPS- or LBP-dependent binding of FITC-LPS to monocytes that was CD14 dependent at up to 100 ng of LPS per ml, and saturation of binding was shown.

Troelstra, A; Antal-Szalmas, P; de Graaf-Miltenburg, L A; Weersink, A J; Verhoef, J; Van Kessel, K P; Van Strijp, J A



Peripheral Fluorescein Angiographic Findings in Fellow Eyes of Patients with Branch Retinal Vein Occlusion  

PubMed Central

Introduction. Branch retinal vein occlusion (BRVO) is a common retinal vascular condition that results in intraocular inflammatory changes. Ultra wide field fluorescein angiography (UWFFA) is a retinal imaging device that can capture peripheral retinal findings. The purpose of this study was to look for peripheral findings in the fellow eye of patients with BRVO using UWFFA. Methods. Retrospective imaging review of patients diagnosed with BRVO that had both eyes imaged with UWFFA. Images were graded for peripheral findings in other quadrants of the same eye as well as in all quadrants of the fellow eye. Results. Of 81 patients, 14 (17%) patients had late vascular leakage in a quadrant other than the BRVO distribution. Five (6%) findings were in the same eye, 8 (10%) findings were in the fellow eye, and 1 (1%) finding was in both the same eye and the fellow eye. Of these 14 patients, 11 (80%) patients had hypertension. Conclusion. Late peripheral retinal leakage in the fellow eye of patients with BRVO was detected in this cohort of patients with UWFFA. This novel finding may represent underlying systemic inflammation, hypertension, or bilateral BRVOs.

Franco-Cardenas, Valentina; Pan, Carolyn K.; Kim, Hanna Y.; Schwartz, Steven D.



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

PubMed Central

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



The analytical measurement of fluorescein, quinine and trace metal concentrations in solution using single bubble sonoluminescence  

NASA Astrophysics Data System (ADS)

A single bubble was generated and levitated in a high-intensity sound field within a spherical flask excited in its fundamental mode. Under optimum experimental conditions the bubble was observed to emit light in the form of short flashes. This phenomenon is known as single bubble sonoluminescence (SBSL). Using this process, the emitted light from the bubble was monitored when solutions containing fluorescein, quinine and sodium, potassium and copper salts were placed in the cell. The results obtained indicated that reproducible signals related directly to the concentration of the species present in solution could be achieved using single bubble sonoluminescence. The results for the molecular species were compared with those obtained by fluorescence spectroscopy and, in the case of quinine, parallel determinations of concentration in a test solution were performed with consistent results. SBSL signals were also observed to exhibit a linear correlation with the concentration of several trace metal salts introduced to the solution in the measurement cell. However, it was not possible to demonstrate that the SBSL signals were derived from stimulated atomic emission or fluorescence, and it was concluded that the effect may result from an indirect effect involving the bubble excitation mechanism.

Wallace, P.; McCallum, K.; Barnard, C. L. R.; Clement, C.; Marshall, J.; Carroll, J.



Evaluation of circulation disorder in coronary slow flow by fundus fluorescein angiography.  


Coronary slow flow (CSF) may be a reflection of a systemic slow-flow phenomenon in the coronary arterial tree. In this study, the CSF group consisted of 24 men (77.4%) and 7 women (22.5%). An age- and gender-matched normal coronary artery (control) group was composed of 21 men (72.4%) and 8 women (27.5%). Retinal arteriovenous circulation time was measured using fundus fluorescein angiography as a part of the microcirculation and the circulation time between the antecubital vein and the retina as a part of the systemic circulation in patients with CSF and controls with normal coronary arteries. The mean arm-retina circulation time was 19.0 ± 5.7 seconds in the CSF group and 14.1 ± 3.1 seconds in the control group (p <0.001). The mean retinal arteriovenous passage time was 2.6 ± 0.9 seconds in the CSF group and 2.1 ± 0.7 seconds in the control group (p = 0.001). Strikingly, retinal findings of chronic central serous retinopathy were observed in 3 patients in the CSF group. In conclusion, CSF may indeed be a part of a systemic slow-flow phenomenon. The association of central serous retinopathy with this condition suggests that corticosteroids and the sympathetic system may play important roles in the pathogenesis of the disease by causing or contributing to increases in microvascular resistance and tonus. PMID:23538021

Koç, Sahbender; Ozin, Bulent; Alt?n, Cihan; Altan Yayc?o?lu, Rana; Ayd?nalp, Alp; Müderrisoglu, Haldun



Engineered single-chain dimeric streptavidins with an unexpected strong preference for biotin-4-fluorescein  

PubMed Central

Streptavidin, a homotetrameric protein with extremely tight biotin binding (Kd ? 10-14 M), has been widely used as an affinity reagent. Its utility would be increased by engineering single-chain mutants with a wide spectrum of affinities, more suitable for phage-display and chip technologies. By a circular permutation procedure, we converted streptavidin to a single-chain dimer (SCD) with two biotin-binding sites and introduced random mutations by error-prone PCR. Clones from a phagemid library, expressed as gene-3 fusion proteins on M13 bacteriophage, were panned with biotinylated beads, and SCD genes from affinity-enriched phage were subcloned to produce soluble proteins. Purification of products from the original gene and two mutants by FPLC and analysis by MALDI-TOF MS showed they exist in both dimeric (single-chain) and tetrameric (two-chain) forms, which were further characterized for their binding affinity to biotin-4-fluorescein (B4F) by fluorescence polarization and intensity measurements. Kd? values for B4F ranged from ?10-11 to 10-10 M, although Kd values for biotin ranged from 10-6 to 10-5 M. These results point to the possibility of combining an SCD streptavidin mutant with B4F derivatives to create a fluorescence-tagged affinity system with tight but still-reversible interaction that could be used sequentially with ordinary streptavidin–biotin for composite separation or analysis steps.

Aslan, Filiz M.; Yu, Yong; Mohr, Scott C.; Cantor, Charles R.



[Purtscher-like retinopathy in acute alcoholic pancreatitis: fluorescein angiography and optical coherence tomography findings].  


27 year old patient with a history of alcohol abuse after consumption of fat meal and wine following epigastric pain noticed sudden bilateral visual loss: right eye logMAR 0, 94, left eye logMAR 1, 22. Retinal examination revealed massive edema in the central part of the retina, multiple cotton wool spots in the posterior pole peripapillary and flame-shaped hemorrhages. On the OCT, there was edema most of all in the inner part of the retina, but also subretinal, hyperreflectivity in the nerve fibre layers corresponding to massive cotton wool spots. Fluorescein angiography in the early phases showed hypofluorescent ischemic areas of the retina subsequently leakage developed in the late phases. Immediately after antibiotic and spasmoanalgetic treatment of the pancreatitis visual acuity improved and 2 months after beginning of the therapy visual acuity is logMAR 0 bilateraly. We proposed that the most suspected cause of Purtscher-like retinopathy in this case is fat embolism. PMID:21394969

Stefanicková, J; Hasa, J; Hlinstáková, S; Pesko, K; Krahulec, B; Strmen, P



A modified fixed staining method for the simultaneous measurement of reactive oxygen species and oxidative responses.  


The generation of reactive oxygen species (ROS) in a live-cell system is routinely measured using the oxidation-sensitive fluorescent probe dichlorofluorescein (DCF). However, it is difficult to simultaneously monitor cellular oxidative responses and ROS generation in cells, and analyses of cellular oxidative responses are typically performed after ROS generation has been evaluated. In this study, we developed a modified fixed staining method that allows the simultaneous analysis of ROS generation and oxidative responses using standard immunostaining techniques. A microplate reader-based assay showed that of the fixatives tested, only methanol did not alter the hydrogen peroxide (H(2)O(2))-mediated oxidation of the responsive dye 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (CM-H(2)DCFDA), a chloromethyl derivative of H(2)DCFDA, or the fluorescence of oxidized DCF in vitro. Further in vivo assays using flow cytometry showed that both methanol and acetic acid maintained the fluorescence of oxidized DCF in H(2)O(2)-, antimycin A-, and serum starvation-treated human lung adenocarcinoma A549 cells and human microvascular endothelial HMEC-1 cells. Following acetic acid-based fixation, the ROS generation in starved HMEC-1 cells could be evaluated by flow cytometric analysis while simultaneously monitoring the phosphorylation status of p38 mitogen-activated protein kinase. Immunostaining also revealed the synchronization of ROS generation and the H(2)O(2)-induced phosphorylation of Src homology-2 domain-containing phosphatase2. This study describes a modified method that may be used in future biomedical investigations to simultaneously measure intracellular ROS production and cellular oxidative responses. PMID:23178299

Shen, Wan-Jou; Hsieh, Chia-Yuan; Chen, Chia-Ling; Yang, Kao-Chi; Ma, Ching-Ting; Choi, Pui-Ching; Lin, Chiou-Feng



Strategic Tool Use: Pencil Box Staining  

NSDL National Science Digital Library

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

Boston, Wghb



Detection of Legionella pneumophila in environmental water samples using a fluorescein conjugated monoclonal antibody.  

PubMed Central

Sixty-three environmental water samples from various sources were examined for the presence of Legionella pneumophila with a commercially available direct fluorescent monoclonal antibody (GS), an indirect fluorescent antibody test (IFAT) and culture. GS detected L. pneumophila in 94% and 100% of environmental water samples which were culture and IFAT positive for L. pneumophila, respectively. IFAT detected 69% of L. pneumophila culture positive samples. Cultures of L. pneumophila serogroups 1 to 12, 14 and non-L. pneumophila bacteria which may be found in water, and bacteria containing non-specific binding proteins, were stained by GS and IFAT. GS identified all serogroups of L. pneumophila and did not cross react with any non-L. pneumophila bacteria. L. pneumophila in environmental samples was easy to detect against a clear dark background when stained with GS. Images Fig. 1

Makin, T.; Hart, C. A.



Effects of sodium citrate plus sodium diacetate and buffered vinegar on Escherichia coli O157:H7 and psychrotrophic bacteria in brine-injected beef.  


The objective of this research was to examine the effects of sodium citrate plus sodium diacetate or buffered vinegar on Escherichia coli O157:H7 and psychrotrophic bacteria when incorporated in brine solutions for injected beef. Two experiments were conducted in which 30 top rounds and 30 top sirloins were injected (110%) to contain (i) 0.5% sodium chloride and 0.4% sodium tripolyphosphate as the control (CNT); (ii) CNT with a 1% solution of 80% sodium citrate plus 20% sodium diacetate (SC + D); or (iii) CNT with 2% buffered vinegar (VIN) in the final product. For the E. coli challenge, muscles were surface inoculated to target 6 log CFU/cm(2). After injection and 10 days of storage in a vacuum package (4°C), one half of each muscle was sampled raw and the other half was cooked to an internal temperature of 60°C with a 12-min hold. For raw samples, a significant reduction of 0.6 and 1.0 log CFU/g of E. coli O157:H7 was observed in both SC + D- and VIN-injected top rounds and sirloins, respectively. All cooked samples were E. coli O157:H7 negative. For psychrotrophic analysis, subprimals were injected and vacuum packaged for 10 days at 0 ± 1°C. After 10 days of storage, steaks were fabricated and placed in aerobic display (4 ± 1°C) for 1, 7, 14, and 21 days. Psychrotrophic organism growth was restricted in SC + D and VIN samples when compared with CNT on all days except day 1. Sodium citrate plus sodium diacetate or buffered vinegar may improve the safety and shelf life of multineedle brine-injected beef. PMID:21375870

Ponrajan, Amudhan; Harrison, Mark A; Segers, Jacob R; Lowe, Bradley K; McKeith, Russell O; Pringle, T Dean; Martino, Karina G; Mulligan, Jake H; Stelzleni, Alexander M



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

Microsoft Academic Search

PEGylated PAMAM conjugated fluorescein-doped magnetic silica nanoparticles (PEGylated PFMSNs) have been synthesized for evaluating their ability across the blood–brain barrier (BBB) and distribution in rat brain. The obtained nanoparticles were characterized by transmission electron microscopy (TEM), thermal gravimetry analyses (TGA), zeta potential (?-potential) titration, and X-ray photoelectron spectroscopy (XPS). The BBB penetration and distribution of PEGylated PFMSNs and FMSNs in

Shuting Ku; Feng Yan; Ying Wang; Yilin Sun; Nan Yang; Ling Ye



A fluorescein tracer release experiment in the hydrothermally active crater of Vailulu'u volcano, Samoa  

NASA Astrophysics Data System (ADS)

On 3 April 2001, a 20 kg point source of fluorescein dye was released 30 m above the bottom of the active summit caldera of Vailulu'u submarine volcano, Samoa. Vailulu'u crater is 2000 m wide and at water depths of 600-1000 m, with the bottom 200 m completely enclosed; it thus provides an ideal site to study the hydrodynamics of an active hydrothermal system. The magmatically driven hydrothermal system in the crater is currently exporting massive amounts of particulates, manganese, and helium. The dispersal of the dye was tracked for 4 days with a fluorimeter in tow-yo mode from the U.S. Coast Guard icebreaker Polar Sea. Lateral dispersion of the dye ranged from 80 to 500 m d-1; vertical dispersion had two components: a diapycnal diffusivity component averaging 21 cm2 s-1, and an advective component averaging 0.025 cm s-1. These measurements constrain the mass export of water from the crater during this period to be 8-1.3+4.6 × 107 m3 d-1, which leads to a "turnover" time for water in the crater of ˜3.2 days. Coupled with temperature data from CTD profiles and Mn analyses of water samples, the power output from the crater is 610-100+350 MW, and the manganese export flux is ˜240 kg d-1. The Mn/Heat ratio of 4.7 ng J-1 is significantly lower than ratios characteristic of hot smokers and diffuse hydrothermal flows on mid-ocean ridges and points to phase separation processes in this relatively shallow hydrothermal system.

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



Medium effects on the prototropic equilibria of fluorescein fluoro derivatives in true and organized solution.  


The stepwise ionization (H(3)R(+) <==> H(2)R <==> HR(-) <==> R(2-)) of four fluorescein fluoro derivatives was studied by visible spectroscopy. The pK(a) values were determined in water, in 50 mass % aqueous ethanol, in oil-in-water microemulsions (benzene + CTAB + pentanol-1 in water with 1.0 M KCl; CTAB = cetyltrimethylammonium bromide), and in reversed ones (water + AOT in n-octane; AOT = bis-2-ethylhexylsulphosuccinate or Aerosol OT). The medium effects, DeltapK(a), i.e., changes in pK(a) of these dyes on going from water to some other solvent systems, were rationalized by considering the tautomerism, the values of microscopic ionization constants, and the charge types of the acid-base couples. An expressed shift of the tautomeric equilibria of H(2)R toward colorless lactone was registered on going from water to both aqueous ethanol and organized solutions. While the monoanions HR(-) of 3',4',5',6'-tetrafluoro- and 2,7,3',4',5',6'-hexafluorofluorescein exist in all the systems studied as a tautomer with ionized carboxylic and nonionized hydroxy groups, in the case of 2,4,5,7-tetrafluorofluorescein, the prevalence of another tautomer was observed (COOH and O(-) groups). For 2,7-difluorofluorescein (Oregon Green 488), the partial shift of the tautomeric equilibrium of HR(-) was registered from (COO(-) and OH) in water to (COOH and O(-)) in other solvent systems. The data for the dyes located in an AOT-based pseudophase indicate that the interior of the latter exerts essential differentiation of the acid strength of the dyes, probably caused by the peculiarity of dye species location in water pools. While the state of tautomeric equilibria resembles that in nonaqueous media, the absorption maxima of R(2-) species are close to those in water. Such nonuniform influence displayed by AOT-based water droplets should be taken into account when examining them by using different molecular probes. PMID:20232888

McHedlov-Petrossyan, Nikolay O; Vodolazkaya, Natalya A; Gurina, Yuliya A; Sun, Wei-Chuan; Gee, Kyle R



Hematoxylin-lac-curcuma polychrome stain for mucin.  


A polychrome method for detection of mucin substance in paraffin section is produced by sequential stepwise staining of hematoxylin, crude lac extract (Laccifer lacca), and crude curcuma extract (khamin shan-Curcuma longa). The name LacCur stain is proposed. After a tissue section is deparaffinized and rehydrated, it is stained with Weigert's hematoxylin for 7 minutes. After a quick wash, it is stained for at least 3 hours with lac dye mordanted with aluminum chloride. Washed again and premordanted with ferric chloride for 1 minute, in the last step, it is counterstained with curcuma dye for 5 minutes. With this staining method, the nuclei are stained black, mucin deep red, and organelles and ground substances brownish yellow. The method and outcome colors are comparable to the widely used Mayer's mucicarmine staining method. It costs less than the Mayer's mucicarmine staining method and the procedure is not complicated. PMID:7543924

Sriplung, H; Kietthubthew, S; Boonyaphiphat, P



21 CFR 864.1850 - Dye and chemical solution stains.  

Code of Federal Regulations, 2013 CFR

...CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical solution stains...hematology. (b) Classification. Class I (general controls). These devices are exempt from the premarket...



Basic blue 93. An alternative to the traditional myeloperoxidase stain.  


This report describes the selective staining of lysosomes in cells of neutrophilic granulocytic origin by the azo textile dye basic blue 93. Used as a single agent aqueous stain after fixation, the dye stained lysosomes black. Stained in this way, lysosomes were sharply delimited and easily visualized. Large numbers of black-stained lysosomes occurred in immature granulocytes like promyelocytes and myelocytes. In more mature granulocytes like metamyelocytes, bands, and neutrophils, substantially fewer numbers of lysosomes were detected. In leukemic myeloblasts and promyelocytes stained with basic blue 93, variable numbers of black-colored lysosomes could be detected and usually paralleled the reactions for myeloperoxidase and Sudan black B in the same cells. As an alternative to the traditional stains for myeloperoxidase, basic blue 93 has several advantages, including simplicity of use, ability to stain aged specimens, and stability of the reaction product. PMID:2449070

Kass, L



BAM R16: Crystal Violet Stain (for Bacteria)  

Center for Food Safety and Applied Nutrition (CFSAN)

... BAM R16: Crystal Violet Stain (for Bacteria). January 2001. Bacteriological Analytical Manual. R16 Crystal Violet Stain (for Bacteria). ... More results from


Polychromatic staining of plant cell walls by toluidine blue O  

Microsoft Academic Search

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

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



Giemsa staining and the distribution of heterochromatin in rye chromosomes  

Microsoft Academic Search

C-banding, by Giemsa staining, is largely restricted to distal regions of rye chromosomes. This applies, also, to B chromosomes. The distribution of C-bands coincides with that of distally localised heterochromatin. The area of metaphase chromosomes staining with Giemsa corresponds to the area occupied by heterochromatin in interphase nuclei as revealed by Feulgen staining. The Giemsa-stained C-bands account, therefore, for all

S C Verma; H Rees



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.

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



Characterization of DNA chips by nanogold staining.  


DNA microarray is an important tool in biomedical research. Up to now, there are no chips that can allow both quality analysis and hybridization using the same chip. It is risky to draw conclusions from results of different chips if there is no knowledge of the quality of the chips before hybridization. In this article, we report a colorimetric method to do quality control on an array. The quality analysis of probe spots can be obtained by using gold nanoparticles with positive charges to label DNA through electrostatic attraction. The probe spots can also be detected by a simple personal computer scanner. Gold nanoparticles deposited on a glass surface can be dissolved in bromine-bromide solution. The same microarray treated with gold particles staining and destaining can still be used for hybridization with nearly the same efficiency. This approach makes quality control of a microarray chip feasible and should be a valuable tool for biomarker discovery in the future. PMID:19328767

Hsiao, Chen-Ren; Chen, Chung-Hsuan



Differential staining of interspecific chromosomes in somatic cell hybrids by alkaline Giemsa stain  

Microsoft Academic Search

Staining of chromosome preparations of Chinese hamster-human hybrid cells and mouse-chimpanzee hybrids with alkaline Giemsa has yielded color differentiation of the interspecific chromosomes. Bicolor chromosomes, indicating apparent translocations also are observed for each of these hybrids. The specific color differences observed provide a rapid means of recognizing and aiding in the identification of the interspecific chromosomes and apparent translocations in

K. K. Friend; S. Chen; F. H. Ruddle



Lignin Staining. A Limited Success in Identifying Koa Growth Rings.  

National Technical Information Service (NTIS)

Among the lignin stains tested in trying to identify growth rings in koa(Acacia koa Gray), phloroglucinol was the most effective. The light colored sapwood of mature trees stained readily, with growth rings apparent. But staining failed to emphasize rings...

H. L. Wick



Cigarette staining and cleaning of a maxillofacial silicone  

SciTech Connect

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

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



Staining methods applied to glycol methacrylate embedded tissue sections  

Microsoft Academic Search

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

P. S Cerri; E Sasso-Cerri



Control of the Staining Procedure after Paper Electrophoresis  

Microsoft Academic Search

WHEN quantitative techniques are attempted for the analysis of serum proteins separated by electrophoresis on filter paper, it is recognized that the staining procedure, followed by removing the surplus stain, constitutes sources of error. In order to get more satisfactory reproducibility we apply on each paper strip (after electrophoresis but before staining) 0.02 ml. of a 0.05 per cent solution

Ch. Wunderly



New selective Giemsa technique for human chromosomes, Cd staining  

Microsoft Academic Search

AFTER the introduction of the quinacrine fluorescence method1, several Giemsa staining techniques have been developed for karyotype analysis of human chromosomes. Pardue and Gall2, originally noticed a denser staining of centromeric regions of chromosomes after in situ hybridisation of mouse chromosome preparations with mouse satellite DNA followed by Giemsa staining. This initial approach was modified by Arrighi and Hsu3 who

Hans Eiberg



7 CFR 3201.87 - Wood and concrete stains.  

Code of Federal Regulations, 2013 CFR

...2013-01-01 false Wood and concrete stains. 3201.87 Section 3201.87 Agriculture...Items § 3201.87 Wood and concrete stains. (a) Definition. Products...qualifying biobased wood and concrete stains. By that date, Federal agencies...



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



Indocyanine green selectively stains the internal limiting membrane  

Microsoft Academic Search

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

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



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



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

National Technical Information Service (NTIS)

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

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



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


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



Investigation on the interaction of tetrachloride fluorescein–bovine serum albumin-?-cyclodextrin and the determination of protein by flow injection analysis  

Microsoft Academic Search

In this paper, a simple and sensitive flow injection analysis (FIA) for the determination of protein with spectroscopic probe was developed. This method was based on the investigation of the interaction of tetrachloride fluorescein (2,4,5,7-tetrachloro-3,6-fluorandiol)–bovine serum albumin (BSA), the coupling reaction of protein with tetrachloride fluorescein (TCFS) which was used as a spectroscopic probe in the presence of ?-cyclodextrin (?-CD).

Xiashi Zhu; Yanyan Hu; Aiqin Gong



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

PubMed Central

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

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



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

Microsoft Academic Search

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

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



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

PubMed Central

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

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



In vivo Delivery of Fluoresceinated Dextrans to the Murine Growth Plate: Imaging of Three Vascular Routes by Multiphoton Microscopy  

PubMed Central

Bone elongation by endochondral ossification occurs through the differentiation cascade of chondrocytes of cartilaginous growth plates. Molecules from the systemic vasculature reach the growth plate from three different directions: epiphyseal, metaphyseal, and via a ring vessel and plexus associated with the perichondrium. This study is an analysis of the real-time dynamics of entrance of fluoresceinated tracers of different molecular weights into the growth plate from the systemic vasculature, and tests the hypothesis that molecular weight is a key variable in the determination of both the directionality and the extent of tracer movement into the growth plate. Multiphoton microscopy was used for direct in vivo imaging of the murine proximal tibial growth plate in anesthetized 4-5-week-old transgenic mice with green fluorescent protein linked to the collagen II promoter. Mice were given an intracardiac injection of either fluorescein (332.3 Da), or fluoresceinated dextrans of 3, 10, 40, 70 kDa, singly or sequentially. For each tracer, directionality and rate of arrival, together with extent of movement within the growth plate, were imaged in real time. For small molecules (up to 10 kDa) vascular access from all three directions was observed and entrance was equally permissive from the metaphyseal and the epiphyseal sides. Within our detection limit (a few per cent of vascular concentration) 40 kDa and larger dextrans did not enter. These results have implications both for understanding systemic and paracrine regulation of growth plate chondrocytic differentiation, as well as variables associated with effective drug delivery to growth plate chondrocytes.

Farnum, Cornelia; Lenox, Michelle; Zipfel, Warren; Horton, William; Williams, Rebecca



Ultrafast electron transfer in the complex between fluorescein and a cognate engineered lipocalin protein, a so-called anticalin.  


Anticalins are a novel class of engineered ligand-binding proteins with tailored specificities derived from the lipocalin scaffold. The anticalin FluA complexes fluorescein as ligand with high affinity, and it effects almost complete quenching of its steady-state fluorescence. To study the underlying mechanism, we have applied femtosecond absorption spectroscopy, which revealed excited-state electron transfer within the FluA*Fl complex to be responsible for the strong fluorescence quenching. On the basis of a comparison of redox potentials, either tryptophan or tyrosine may serve as electron donor to the bound fluorescein group in its excited singlet state, thus forming the fluorescein trianion radical within 400 fs. The almost monoexponential rate points to a single, well-defined binding site, and its temperature independence suggests an (almost) activationless process. Applying conventional electron transfer theory to the ultrafast forward and slower back-rates, the resulting electronic interaction is rather large, with approximately 140 cm(-1) for tyrosine, which would be consistent with a coplanar arrangement of both aromatic moieties within van der Waals distance. The weak residual steady-state fluorescence originates from a small (approximately 10%) component with a time constant in the 40-60 ps range. These results demonstrate the power of time-resolved absorption spectroscopy as a diagnostic tool for the elucidation of a fluorescence quenching mechanism and the temporal profiles of the processes involved. The high structural and dynamic definition of the complexation site suggests the anticalin FluA to be a promising model in order to tailor and probe electronic interactions and energetics in proteins. PMID:11900559

Götz, M; Hess, S; Beste, G; Skerra, A; Michel-Beyerle, M E



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


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

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



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.



Effect of Sodium Fluorescein and Plating Medium on Recovery of Irradiated Escherichia coli and Serratia marcescens from Aerosols  

PubMed Central

Irradiation of aerosols of either Escherichia coli or Serratia marcescens with simulated solar (xenon) radiation caused a significant decrease in viability. When sodium fluorescein was employed to determine the physical loss of organisms from the aerosol, an additional adverse effect upon survival was noted. The decay curves indicated that at least two mechanisms of inactivation were operative, one due to aerosolization, the other to irradiation. After collection from aerosols, both species of microorganisms grew better on blood agar base than on Casitone agar, but this finding did not appear to be related to the effect of irradiation.

Dorsey, Emerson L.; Berendt, Richard F.; Neff, Everett L.



Involvement of the eye in SLE and scleroderma. A study using fluorescein angiography in addition to clinical ophthalmic assessment.  

PubMed Central

General examination of the eye was carried out in 22 patients with systemic lupus erythematosus (SLE) and in 10 with scleroderma. 3 of the SLE and 2 of the scleroderma patients had keratoconjunctivitis sicca. Fluorescein angiography showed abnormalities of the retinal vasculature in one of a subgroup of 12 SLE patients and one of 10 scleroderma patients. None of the 12 SLE patients had abnormalities of the choroidal vasculature, while 5 of the 10 scleroderma patients had patchy areas of nonperfusion of the choroidal capillary bed. Images

Grennan, D M; Forrester, J



Harmonization of the intracellular cytokine staining assay.  


Active immunotherapy for cancer is an accepted treatment modality aiming to reinforce the T-cell response to cancer. T-cell reactivity is measured by various assays and used to guide the clinical development of immunotherapeutics. However, data obtained across different institutions may vary substantially making comparative conclusions difficult. The Cancer Immunotherapy Immunoguiding Program organizes proficiency panels to identify key parameters influencing the outcome of commonly used T-cell assays followed by harmonization. Our successes with IFN?-ELISPOT and peptide HLA multimer analysis have led to the current study on intracellular cytokine staining (ICS). We report the results of three successive panels evaluating this assay. At the beginning, 3 out of 9 participants (33 %) were able to detect >6 out of 8 known virus-specific T-cell responses in peripheral blood of healthy individuals. This increased to 50 % of the laboratories in the second phase. The reported percentages of cytokine-producing T cells by the different laboratories were highly variable with coefficients of variation well over 60 %. Variability could partially be explained by protocol-related differences in background cytokine production leading to sub-optimal signal-to-noise ratios. The large number of protocol variables prohibited identification of prime guidelines to harmonize the assays. In addition, the gating strategy used to identify reactive T cells had a major impact on assay outcome. Subsequent harmonization of the gating strategy considerably reduced the variability within the group of participants. In conclusion, we propose that first basic guidelines should be applied for gating in ICS experiments before harmonizing assay protocol variables. PMID:22714399

Welters, Marij J P; Gouttefangeas, Cécile; Ramwadhdoebe, Tamara H; Letsch, Anne; Ottensmeier, Christian H; Britten, Cedrik M; van der Burg, Sjoerd H



Differences between the peripheral and the central nervous system in permeability to sodium fluorescein.  


Sodium fluorescein (SF) was used as a very small tracer (mol wt 376; 5 A diameter) to examine diffusion barriers in peripheral nerves and to compare them to those in other regions of the nervous system. The technique involved immobilization of the tracer by rapid freezing, followed by freeze-drying and vacuum embedding in paraffin. The localization of the SF was then determined in tissue secretions using fluorescence microscopy. Even at the highest doses of intravenously (IV) injected tracer, no extravasation could be detected in the cerebral cortex. On the other hand, SF penetrated very rapidly into peripheral ganglia and into the epineurium and perineurium of large peripheral nerves. The penetration of SF into the endoneurium of large nerves was, however, much more restricted with tracer detectable within the endoneurium only at high doses and long survival times. Even in such cases, the level of SF fluorescence was much lower within nerve fascicles than in the epineurium and the perineurium, and a sharp gradient in fluorescence intensity persisted at the inner border of the perineurium. The extent of extravasation into the endoneurium varied markedly betwen different fascicles of the same nerve and between different nerves in the same animal. Experiments involving injection of high doses of SF adjacent to the nerve indicated relatively little movement of SF across the perineurium, which indicates that the observed accumulation of tracer within the endoneurium was the result of direct extravasation of SF from the endoneural blood vessels. Small nerve branches (< 100 mu diameter) showed an earlier and more extensive penetration of SF into the endoneurium than large nerves like the sciatic, hypoglossal, or ventral tail nerve. This may be due to a diffusion of SF along the extracellular space of the endoneurium from nerve terminals where the perineurial barrier is open-ended. In experiments involving IV injection of a solution containing both green fluorescent SF and red fluorescent Evans Blue (Evans Blue-serum albumin conplex, EBA = mol wt. 69,000), the distribution of SF could be directly compared at various sites and sacrifice times to that of EBA, a much larger tracer. SF appeared more rapidly and extensively than EBA in the various compartments in ganglia and peripheral nerve. The distribution of EBA was the same as is typically seen when this tracer is injected alone, indicating that there was no change in vascular permeability associated with IV injection of SF. Since SF is of very small size, freely diffusible, nontoxic, and detectable at very low concentrations, it should be a useful complement to existing tracers. When tissues are processed according to the indicated procedure, one can obtain a very sensitive and reliable localization of this tracer which should be of value for studies in the nervous system concerning various pathological conditions associated with permeability alterations. PMID:7400388

Malmgren, L T; Olsson, Y



Identifying different types of chromatin using giemsa staining.  


Mixtures of polychrome methylene blue-eosin Y (i.e., Giemsa stain) are widely used in biological staining. They induce a striking purple coloration of chromatin DNA (the Romanowsky-Giemsa effect), which contrasts with the blue-stained RNA-containing cytoplasm and nucleoli. After specific prestaining treatments that induce chromatin disorganization (giving banded or harlequin chromosomes), Giemsa staining produces a differential coloration, with C- and G-bands appearing in purple whereas remaining chromosome regions are blue. Unsubstituted (TT) and bromo-substituted (BT) DNAs also appear purple and blue, respectively. The same occurs in the case of BT and BB chromatids.In addition to discussing the use of Giemsa stain as a suitable method to reveal specific features of chromosome structure, some molecular processes and models are also described to explain Giemsa staining mechanisms of chromatin. PMID:24162977

Stockert, Juan C; Blázquez-Castro, Alfonso; Horobin, Richard W



Standard Care vs Corticosteroid for Retinal Vein Occlusion (SCORE) Study System for Evaluation of Stereoscopic Color Fundus Photographs and Fluorescein Angiograms  

PubMed Central

Objective To describe the procedures and reproducibility for grading stereoscopic color fundus photographs and fluorescein angiograms of participants in the SCORE Study. Methods Standardized stereoscopic fundus photographs and fluorescein angiograms taken at 84 clinical centers were evaluated by graders at a central reading center. Type of retinal vein occlusion (RVO), area of retinal thickening, and area of retinal hemorrhage are evaluated from fundus photographs; area of fluorescein leakage and area of capillary nonperfusion are measured on fluorescein angiography. Temporal reproducibility consisted of annual regrading of a randomly selected dedicated subset of fundus photographs (60 subjects) and fluorescein angiograms (40 subjects) for 3 successive years. Contemporaneous reproducibility involved monthly regrading of a 5% random selection of recently evaluated fundus photographs (n=73). Results The intergrader agreement for RVO type and presence of retinal thickening was greater than 90% in the 3 annual regrades. The intraclass correlation (ICC) for area of retinal thickening in the 3 years ranged from 0.39 to 0.64 and for area of retinal hemorrhage, 0.87 to 0.96. The ICC for area of fluorescein leakage ranged from 0.66 to 0.75 and for capillary nonperfusion, 0.94 to 0.97. The contemporaneous reproducibility results were similar to those of temporal reproducibility for all variables except area of retinal thickening (ICC, 0.84). Conclusions The fundus photography and fluorescein angiography grading procedures for the SCORE Study are reproducible and can be used for multicenter longitudinal studies of RVO. A systematic temporal drift occurred in evaluating area of retinal thickening.

Blodi, Barbara A.; Domalpally, Amitha; Scott, Ingrid U.; Ip, Michael S.; Oden, Neal L.; Elledge, Julee; Warren, Kelly; Altaweel, Michael M.; Kim, Judy E.; Van Veldhuisen, Paul C.



Techniques for Controlling Variability in Gram Staining of Obligate Anaerobes  

Microsoft Academic Search

Identification of anaerobes recovered from clinical samples is complicated by the fact that certain gram- positive anaerobes routinely stain gram negative;Peptostreptococcus asaccharolyticus,Eubacterium plautii,Clos- tridium ramosum, Clostridium symbiosum, and Clostridium clostridiiforme are among the nonconformists with regard to conventional Gram-staining procedures. Accurate Gram staining of American Type Culture Collec- tion strains of these anaerobic bacteria is possible by implementing fixing and



Differential giemsa staining of sister chromatids after extraction with acids  

Microsoft Academic Search

Chromosomes of Chinese hamster strain cells were air-dried on slides after BrdU substitution for two or three rounds of replication. The preparations were treated with 20% PCA at 55° C for 20–30 min, or 5N HCl at 55° C for 15–20 min. After staining with Giemsa, unifilarly BrdU-substituted chromatids stained faintly and bifilarly substituted chromatids stained darkly. Such a pattern

Susumu Takayama; Shunichi Sakanishi



Factors involved in differential giemsa-staining of sister chromatids  

Microsoft Academic Search

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

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



Victoria blue B — a nuclear stain for cytology  

Microsoft Academic Search

The aim of this study was to investigate the staining characteristics of Victoria Blue B in alcohol solutions. Cytological specimens (liver and spleen tissue imprints, blood smears) were stained with methanol solutions of commercially available Victoria Blue B-Cl and with pure Victoria Blue B-BF4. The dye concentration, staining time, and protone concentration of the dye solution were varied. The dye

E. Schulte; D. Wittekind; V. Kretschmer



Bis(dimethyl-ammonium) 2,2'-(1,3,6,8-tetra-oxo-2,7-diaza-pyrene-2,7-di-yl)diacetate.  


The asymmetric unit of title compound, 2C(2)H(8)N(+)·C(18)H(8)N(2)O(8) (2-), comprises one crystallographically independent dimethyl-ammonium cation and half of a 2,2'-(1,3,6,8-tetra-oxo-2,7-diaza-pyrene-2,7-di-yl)diacetate dianion. The anion lies on an inversion centre and the two carboxyl-ate groups are in trans positions based on the naphthaleneteracarb-oxy-lic diimide group. The crystal packing is stabilized by N-H?O hydrogen bonds between cations and anions, as well as by ?-? inter-actions between the naph-thaleneteracarb-oxy-lic diimide groups [centroid-centroid distance = 4.812?(3)?Å]. PMID:22091009

Xu, Lan-Ping; Zhao, Wen-Na; Han, Lei



Histochemical Staining Methods for Localizing Cyanoacrylate Polymers in Tissue Sections.  

National Technical Information Service (NTIS)

A histochemical staining procedure was devised which makes it possible to visually locate and study clinically implanted polymers of alkyl alpha cyanoacrylates and isobutyl in tissue sections. (Author)

K. C. Pani W. M. McAllister F. Leonard P. M. Margetis



Staining of neuroendocrine cells by Linder's argyrophil method.  


Linder's argyrophil method, recently developed to stain nervous structures, is useful in the histochemical study of amine- and/or peptide-producing neuroendocrine (APUD) cells. On sections from various organs of four animal species Linder's method worked well and rapidly stained the neuroendocrine cells yellow, red or black; it stained black nervous structures against a pale yellow background. Double staining of single sections from Bouin-fixed gastric mucosa of rabbits demonstrated the correspondence of both Linder- and Grimelius-positive cells. Rapidity of application, intensity of impregnation and reproducibility in results are the best features of Linder's method when applied to the study of the neuroendocrine system. PMID:2461924

Cecio, A; Vittoria, A; Budetta, G; Corona, M



A quantitative study of fluorescein isothiocyanate-dextran transport in the microcirculation of the isolated perfused rat liver.  


Hepatic extraction of solutes depends on microvascular angioarchitecture, hemodynamics and solute concentrations. These factors may contribute to the heterogeneity observed in solute transport and uptake in the hepatic lobules. However, predictions of liver extraction based on black-box models require assumptions about these factors and the microvascular transport mechanisms involved. Consequently, the purpose of this study was to investigate solute transport and uptake by hepatocytes. Livers from male Sprague-Dawley rats were perfused at physiological flowrates and portal pressures on the stage of an in vivo microscope using a low-hematocrit Ringer solution. A bolus of fluorescein isothiocyanate-dextrans (17,900, 39,000, 65,600 or 156,900 MW), which are considered inert fluid-phase markers, was injected into the portal vein. Fluorescein isothiocyanate fluorescence, as a measure of solute concentration, was video recorded in periportal or centrivenular regions of the lobules. Spatial and temporal fluorescence data, measured in sinusoids and hepatocytes, were fit to one-dimensional transport models to determine estimates for an intracellular effective diffusion coefficient and for hepatocyte permeability. The calculated effective diffusion coefficients were 2.5 times larger for dextrans less than 66,000 MW, but were not different between the periportal and centrivenular regions. Also, the values did not show the inverse log-log molecular weight dependency for dextrans seen in other microvascular tissues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2461894

Stock, R J; Cilento, E V; McCuskey, R S



Disinfection of seawater for hatchery aquaculture systems using electrolytic water treatment  

Microsoft Academic Search

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

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



Transformed roots of Artemisia annua exhibit an unusual pattern of border cell release  

Microsoft Academic Search

Summary  Border cells from Artemisia annua were examined from hairy roots grown in shake flasks, culture plates, a bubble column reactor, and a nutrient mist (aeroponic)\\u000a reactor. When well-hydrated roots were subjected to shear, border cells were first released as an agglomerate and did not\\u000a disperse for several hours. Staining with neutral red and fluorescein diacetate (FDA) showed that both agglomerates

Pamela J. Weathers; Yoo Jeong Kim



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




Microsoft Academic Search

e reviewed 194 revision arthroplasties of the hip and knee performed over a ten-year period. The results of intraoperative Gram staining were available in 169 (87%). Thirty-two were found to be infected (11 hips and 21 knees) and 137 had no evidence of infection. Intraoperative Gram staining was negative in all 169 cases. The method therefore had a sensitivity of



DNA Staining in Agarose Gels with ZN-Cyclen-Pyrene  

Microsoft Academic Search

A pyrene-labeled Zn-cyclen complex for the staining of DNA in agarose gels is reported. The metal chelate coordinates reversibly to the DNA phosphate backbone, which induces the formation of pyrene excimers. The typical pyrene excimer emission is used for the detection of the DNA. Staining is limited to agarose gels and is less sensitive than ethidium bromide, but DNA amounts

Florian Schmidt; Jennifer Schmidt; Alexander Riechers; Susann Haase; Anja-Katrin Bosserhoff; Jörg Heilmann; Burkhard König



Sudan Black Method for Staining Sites of Calcification  

Microsoft Academic Search

IN 19581 a method was described for staining sites of calcification at the `calcification front'-the edge of the preosseous matrix in bone, the cartilage around the hypertrophic cells of the epiphyseal cartilage and the junction of predentin arid dentin. The method consisted, after fixation, in extracting the tissue with hot pyridine, decalcifying, embedding in gelatin, and staining with an alcoholic

J. T. Irving



Investigation of Methods for Removing Stains from Mortar and Concrete.  

National Technical Information Service (NTIS)

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

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



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



Selection and Application of Exterior Stains for Wood.  

National Technical Information Service (NTIS)

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

R. S. Williams W. C. Feist



Removing foxing stains from old paper at 157nm  

Microsoft Academic Search

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

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



Histological staining as a measure of stress in collagen fibers.  


It has been reported previously that collagen fibers will stain either red or green by Masson's and other trichrome methods depending on whether they have been respectively stressed or relaxed prior to fixation. This was shown in skin [1, 2, 3] tendon [4, 5] bone [6] and films of collagen [7]. If this stain-stress dependence is of a unique quantitative nature, then staining could be used as a tension probe for collagen fibers. Relaxed and stressed collagen bundles of rat tail tendon and rat Achilles tendon have been stained using various staining periods, and results indicate that the change in staining may be associated with denser packing of the fibers in the bundle under stress rather than directly due to the stress itself. Denser packing may reduce the rate of penetration of the counterstain thus causing the staining differences. Since this rate of penetration is dependent on a number of other variables (unrelated to stress), it is concluded that collagen staining is not a reliable tension probe. PMID:6204124

Lanir, Y; Walsh, J; Soutas-Little, R W



Stain reduction of an integrated oral hygiene system.  


This article discusses research to determine the efficacy of a prototype integrated power toothbrush and toothpaste dispensing system, the IntelliClean System from Sonicare and Crest, in the removal of extrinsic stain. The prototype integrated system and a positive control, the Sonicare Elite with conventional toothpaste, were evaluated in 2 randomized, single-blinded, parallel 4-week controlled clinical trials. There was a low dropout rate, with 28 subjects of the 31 randomized in study 1 completing the study (10% loss to follow-up) and 26 subjects of the 28 randomized in study 2 completing the study (7% loss to follow-up). Lobene stain scores were used to assess the extent and intensity of stain for all teeth meeting the criteria for inclusion in the studies. Lobene stain scores were assessed at baseline and after 4 weeks in both studies. A survey also was conducted at the conclusion of each study to determine user attitude toward the integrated system. The prototype integrated system was found to significantly reduce overall extrinsic stain over time, performing not significantly differently from the positive control. Overall, the prototype integrated system reduced the composite measure of stain that encompasses both the extent and intensity of stain by 60%. This research demonstrates that the IntelliClean System from Sonicare and Crest is highly effective in reducing extrinsic stain. PMID:15637979

Nunn, Martha E; Chaves, Eros S; Gallagher, Andrew C; Rodriguez, Sally M; Ortblad, Katherine M



[Diagnosis of Chlamydia trachomatis by culture and Macchiavella's staining technique].  


The author modified and tested staining according to Macchiavelli for detection of C. trachomatis in tissue cultures. As compared with IF staining by the monoclonal antibody the author found a high sensitivity (95.7%) and specificity (99.0%) of this method. Giemsa staining evaluated in passing light has a lower sensitivity (85.5%) and lower specificity (94.1%). The advantage of Macchiavelli staining is also the low price of the chemicals used. Imunofluorescent staining with the monoclonal antibody remains also now the reference method and is most reliable. For some materials which are obtained with difficulty (e.g. punctates etc.) the author recommends to use the IF technique. PMID:1724208

Zampachová, E



Some effects of mucopolysaccharide stains on platelet aggregation  

PubMed Central

Five mucopolysaccharide stains inhibit platelet aggregation induced by several aggregating agents: with two stains inhibition is competitive. Alcian blue, 0·05 mg/ml, added to platelet-rich plasma potentiates ADP-induced aggregation. Alcian blue, 0·5 mg/ml, itself produces platelet aggregation, probably causing the release reaction. Three of these stains produce aggregation of red cells. They also bind heparin, and so may influence heparin-neutralizing sites (platelet factor 4) and probably the charge on the platelet membrane. The only known common factor to these stains is their ability to bind onto the mucopoly-saccharides. It is suggested that when platelets are `stained' the surface mucopolysaccharide is altered and that this alteration can influence platelet aggregation.

O'Brien, J. R.



Dynamic staining of bacteria at a single-cell level  

NASA Astrophysics Data System (ADS)

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

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



CGP stain: An inexpensive, odorless, rapid, sensitive, and in principle in vitro methylation-free Coomassie Brilliant Blue stain  

Microsoft Academic Search

Coomassie Brilliant Blue (CBB) protein stains are inexpensive but detect proteins at only at microgram levels. Because of acetic acid and methanol, they cause skin irritation and reduce work motivation by malodor. Recent mass spectrometric (MS) analyses demonstrated that nanogram-sensitive colloidal CBB staining resulted in in vitro methylations of proteins. We propose a rapid, inexpensive, sensitive, odorless, less harsh, and

Hidetaro Yasumitsu; Yasuhiro Ozeki; Sarkar M. A. Kawsar; Tosifusa Toda; Robert Kanaly



Modification of the Mycobacterium bovis extracellular protein MPB70 with fluorescein for rapid detection of specific serum antibodies by fluorescence polarization.  

PubMed Central

The principle of fluorescence polarization described by Perrin (F. Perrin, J. Phys. Radium 7:390-401, 1926) was applied to the development of a novel assay that used fluorescein-labeled Mycobacterium bovis secretory protein MPB70 for rapid detection of anti-MPB70 antibodies in selected sera from three M. bovis-infected species (elk, Ilama, and bison). Labeling of purified MPB70 with fluorescein isothiocyanate resulted in the incorporation of 0.96 +/- 0.08 (mean +/- standard deviation; n = 3) fluorescein group per MPB70 molecule. The labeled protein fluoresced strongly with an emission maximum at 518 nm when excited with light of a wavelength near 493 nm, and its immunoreactivity with anti-MPB70 monoclonal antibody 4C3/17 was not altered by modification with fluorescein. The fluorescence polarization assay protocol was optimized for analysis of serum samples by incorporating into the assay buffer 0.05% lithium dodecyl sulfate, which prevents the occurrence of some nonspecific interactions. Sera from M. bovis-infected animals, selected on the basis of exhibiting the presence of anti-MPB70 antibodies, as detected by enzyme-linked immunosorbent assay (ELISA), reacted with fluorescein-labeled MPB70, resulting in an increase in polarization of up to 330 milli-polarization units, in contrast to the values for noninfected sera (167 to 178 mP), which were close to that obtained in the absence of specific antibodies (164.7 +/- 3.3 mP; n = 6). These results demonstrated the feasibility of using fluorescein-labeled MPB70 to detect anti-MPB70 antibodies by fluorescence polarization and suggested that the assay described here can be an alternative to ELISA or other antibody assay systems. The advantages of this original methodology and its general applicability to the diagnosis of infectious diseases are discussed.

Lin, M; Sugden, E A; Jolley, M E; Stilwell, K



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.

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



Homogeneous luminescent stain etched porous silicon elaborated by a new multi-step stain etching method  

NASA Astrophysics Data System (ADS)

This paper presents a new method to produce porous silicon which derived from the conventional stain etching (SE) method. But instead of one etching step that leads to formation of porous layer, the substrate is subjected to an initial etching step with a duration ?t0 followed by a number of supplementary short steps that differs from a layer to another. The duration of the initial step is just the necessary time to have a homogenous porous layer on the whole surface of the substrate. It was found that this duration is largely dependent of the doping type and level of the silicon substrate. The duration of supplementary steps was kept as short as possible to prevent the formation of bubbles on the silicon surface during silicon dissolution which leads generally to inhomogeneous porous layers. It is found from surface investigation by atomic force microscopy (AFM) that multistep stain etching (MS-SE) method allows to produce homogeneous porous silicon nanostructures compared to the conventional SE method. The chemical composition of the obtained porous layers has been evaluated using Fourier transform infrared spectroscopy (FTIR). Photoluminescence (PL) measurement shows that porous layers produced by SE and MS-SE methods have comparable spectra indicating that those layers are composed of nanocrystallites with comparable sizes. But the intensity of photoluminescence of layer elaborated by MS-SE method is higher than that elaborated by the SE method. Total reflectance characteristics show that the presented method allows the production of porous silicon layers with controllable thicknesses and optical properties. Results for porous silicon layers elaborated on heavily doped n-type silicon show that the reflectance can be reduced to values less than 3% in the major part of the spectrum.

Hajji, M.; Khalifa, M.; Slama, S. Ben; Ezzaouia, H.



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

PubMed Central

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

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



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

NASA Astrophysics Data System (ADS)

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

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



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

PubMed Central

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

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



Simple histochemical stain for acrosomes on sperm from several species.  


The acrosome reaction is an exocytotic process that enables a sperm to penetrate the zona pellucida and fertilize an egg. The process involves the fenestration and vesiculation of the sperm plasma membrane and outer acrosomal membrane releasing the acro somal contents. Many different methods have been devel oped to detect the acrosomal status of sperm. These techniques are sometimes complicated, costly, and can be used on only a few species. The aim of this study was to develop an efficient and inexpensive method to assess the acrosomal status of sperm from a variety of species. We prepared and fixed sperm from humans, cattle, swine, rabbits, guinea pigs, and mice and stained them with Coomassie G250. The acrosomes were stained intensely blue in color. Following capacitation, some sperm were incubated for 1 hr with 10 microM calcium ionophore A23187 to induce the acrosome reaction. They were also stained with Coomassie G-250. Ionophore-treated sperm lacked Coomassie staining over the acrosomal region. Differential interference contrast (DIC), bright field microscopy or Pisum sativum agglutinin staining confirmed that the acrosomes of sperm from these species were reacted in response to calcium ionophore treatment and the acrosome reaction frequencies matched results with Coomassie staining. These results demonstrate that the acrosomal status of mammalian sperm from several species can be determined easily and reliably using this simple Coomassie Blue G-250 staining method. PMID:10092125

Larson, J L; Miller, D J



Improved method for combination of immunocytochemistry and Nissl staining  

PubMed Central

Nissl-staining is a widely used method to study morphology and pathology of neural tissue. After standard immunocytochemistry, the Nissl-staining labels only the nucleus of neurons and the characteristic staining of the neuronal perikarya is absent or very weak. We hypothesized that the RNA degradation during the immunocytochemical treatment results in the loss of cytoplasmic staining with Nissl-dyes. To test this hypothesis, we used RNAse-free conditions for all steps of immunostaining. To further prevent the RNA-degradation by RNAse contaminations, the RNAse inhibitor heparin was added to all antibody-containing solutions. The efficiency of Nissl-staining after standard and RNAse-free double-labeling immunocytochemistry was compared using antibodies against c-Fos and neuropeptide Y (NPY) on tissues of rats refed after three days of fasting. After standard immunocytochemistry, the Nissl-staining labeled the nuclei of neurons and only very faintly the cytoplasm of these cells. The RNAse-free treatment did not alter the distribution of immunoreaction signal, but preserved the staining of neuronal perikarya by the Nissl-dyes. In conclusion, the RNAse-free conditions during immunocytochemistry, allows the labeling of neuronal perikarya by Nissl-dyes. The described method facilitates the mapping of immunocytochemical signals and makes possible the light microscopic examination of the innervation of neurons identified by their nuclear protein content.

Kadar, Andrea; Wittmann, Gabor; Liposits, Zsolt; Fekete, Csaba




PubMed Central

Among the techniques which have been reported to stain the surface coat of cells, for electron microscopy, is lanthanum staining en bloc. Similarly, the presence of the cationic dye, Alcian blue 8GX, in a primary glutaraldehyde fixative has been reported to improve the preservation of the surface coat of cells of many types; however, the preserved coat is not very electron opaque unless thin sections are counterstained. The present paper shows that for several rat tissues lanthanum staining en bloc is an effective electron stain for the cell surface, giving excellent contrast, if combined sequentially with prefixation in an aldehyde fixative containing Alcian blue. The cationic substance cetylpyridinium chloride was found to have a similar effect to that of Alcian blue in enhancing the lanthanum staining of the surface coat material of the brush border of intestinal epithelial cells. The patterns of lanthanum staining obtained for the tissues studied strikingly resemble those reported in the literature where tissues are stained by several standard methods for demonstrating mucosubstances at the ultrastructural level. This fact and the reproduction of the effect of Alcian blue by cetylpyridinium chloride constitute a persuasive empirical argument that the material visualized is a mucopolysaccharide or mucopolysaccharide-protein complex.

Shea, Stephen M.



Passive permeability and outward active transport of fluorescein across the blood-retinal barrier in early ARM  

PubMed Central

AIM—To study the passive and active transport of fluorescein across the blood-retina barrier in early age related maculopathy (ARM) (soft drusen > 63 µm, hyperpigmentation and/or hypopigmentation in patients above 50 years of age).?METHODS—15 patients and 10 healthy subjects were included. Morphological changes were graded from 30 degrees fundus photographs using a simplified version of the epidemiological ARM study group classification system. Differential vitreous spectrofluorophotometry was used to assess the transport properties of the blood-retina barrier (that is, passive permeability and unidirectional permeability caused by outward active transport from the vitreous to the blood).?RESULTS—The passive permeability of the patient group was not significantly different from that of the control group. Four patients with passive permeability more than 3 SD above the mean of the control group (mean 1.8 (SD 0.7) nm/s, range 1.0-3.0 nm/s, data normally distributed) all had centrally located drusen >500 µm and superjacent pigment clumps of 63-500 µm in diameter. There was no significant difference between the unidirectional permeabilities for the patient group and for the control group (mean 47.4 (29.3) nm/s, range 12.7-91.1 nm/s).?CONCLUSION—There was no significant difference in the passive permeability and in the unidirectional permeability of fluorescein. However, the study may indicate that the combination of very large drusen and superjacent pigment clumps in ARM may be associated with a deterioration of the blood-retina barrier.??

Moldow, B.; Larsen, M.; Sander, B.; Lund-Andersen, H.



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.



Basic blue 148: a rapid stain for T helper cells.  


After brief exposure to an aqueous solution of the oxazine textile dye C.I. basic blue 148 following fixation in 37% formalin, 95% ethanol and glacial acetic acid, T helper cell nuclei and cytoplasm in specimens of peripheral blood displayed a deep red-violet color. No other cell in normal blood or bone marrow specimens showed intense staining of this type. The total staining time is 1 min. Basic blue 148 stain is a promising technique for hematology and immunology laboratories as a rapid screening test for T helper cells in blood specimens using a microscopic slide and ordinary incandescent illumination. PMID:7578596

Kass, K



[Characteristics on staining of leukocytes in experimental animals].  


The staining characteristics of the peripheral blood cells from mouse, rat, guinea pig, rabbit, dog, marmoset and monkey were studied. In marmoset, it is easy to distinguish neutrophils from eosinophils by using the phosphate-buffered solution of pH 5 or 6. It was found in the special staining methods that neutrophil granules showed intense peroxidase and Sudan black B reactions in marmoset in comparison with those in the other species of experimental animals. Neutrophil granules rabbit was, however, intensely stained with esterase and acid phosphatase. PMID:1706668

Futamura, Y; Matsumoto, K; Furuya, T



A new technique for Gram staining paraffin-embedded tissue.  

PubMed Central

Five techniques for Gram staining bacteria in paraffin sections were compared on serial sections of pulmonary tissues from eight bacteriological necropsies. Brown and Hopp's method was the most satisfactory for distinguishing Gram-positive and Gram-negative bacteria. However, this method cannot be recommended as the preparations were frequently overstained, and the Gram-negative bacteria were stained indistinctly. A modification of Brown and Hopps' method was developed which stains larger numbers of Gram-negative bacteria and differentiates well between different cell types and connective tissue, and there is no risk of overstaining.

Engbaek, K; Johansen, K S; Jensen, M E



Filtration-based staining of proteins on membranes.  


A filtration-based staining method has been developed for sensitive estimation of proteins in a batch of samples. It is based on focused absorption of an applied protein standard (or sample) and dye solution on nitrocellulose membrane through an aqueous network of capillary channels formed between the membrane and a wetted filter paper. The method does not require any equipment for creating vacuum. Compared with the conventional pouring methods, the new approach reduces the consumption of staining solution and lowers the staining and destaining times (from 1.5 h to approximately 10 min) without compromising the sensitivity. PMID:18482569

Acharya, Debopam; Saha, Debjani; Roy, Dipika; Jain, Priyanka; Dhar, Tarun K



The acid-fast stain is a superior stain for use in the mean mature spermatid count for testicular biopsies.  


The mean mature spermatid count (MMSC) provides a useful, simplified quantitative evaluation of human spermatogenesis that is based on the number of mature spermatids in histological sections of testicular biopsies. Here, the activity of the acid-fast (AF) stain was compared to that of the usual hematoxylin and eosin (H&E) stain in performing the MMSC. Thirty bilateral testicular biopsies showing normal spermatogenesis were chosen retrospectively from 15 subfertile patients with obstructive azoospermia or severe oligospermia. The MMSC was determined on each biopsy by utilizing both H&E and AF stains. The AF stain proved to be specific for the mature spermatids normally counted for the MMSC. It simplified recognition of mature spermatids, thereby shortening the overall time required for the procedure. The mean AF MMSC was lower than the mean H&E MMSC, and the mean interobserver differences were decreased. The AF stain is a superior stain for the MMSC when used in conjunction with the H&E stain for descriptive histology. PMID:9639043

Magid, M S; Ma, Z W; Girardi, S K; Goldstein, M



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



Aneuploidy Test Development: Kinetochore Staining in Mammalian Systems.  

National Technical Information Service (NTIS)

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

R. R. Tice V. L. Dellarco



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

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

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



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

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



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




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

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


The electrical conduction variation in stained carbon nanotubes  

NASA Astrophysics Data System (ADS)

Carbon nanotubes become stained from coupling with foreign molecules, especially from adsorbing gas molecules. The charge exchange, which is due to the orbital hybridization, occurred in the stained carbon nanotube induces electrical dipoles that consequently vary the electrical conduction of the nanotube. We propose a microscopic model to evaluate the electrical current variation produced by the induced electrical dipoles in a stained zigzag carbon nanotube. It is found that stronger orbital hybridization strengths and larger orbital energy differences between the carbon nanotube and the gas molecules help increasing the induced electrical dipole moment. Compared with the stain-free carbon nanotube, the induced electrical dipoles suppress the current in the nanotube. In the carbon nanotubes with induced dipoles the current increases as a result of increasing orbital energy dispersion via stronger hybridization couplings. In particular, at a fixed hybridization coupling, the current increases with the bond length for the donor-carbon nanotube but reversely for the acceptor-carbon nanotube.

Sun, Shih-Jye; Wei Fan, Jun; Lin, Chung-Yi



Interior, detail closeup shot of window with stained glass inserts ...  

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

Interior, detail closeup shot of window with stained glass inserts in top southeast room taken from ther west - J. Weingartner & Son Cigar Factory, 414 East Walnut Street, North Wales, Montgomery County, PA


Digital analysis of mouthrinses' staining characteristics on provisional acrylic resins.  


Provisional restorations are expected to be both aestethically and physically durable during the preparation of permanent restorations. In this study, the staining properties of mouthrinses containing chlorhexidine gluconate, benzydamine hydrochloride and a hybrid mouthrinse were investigated on light and dark shades of a provisional acrylic resin. Totally 80 specimens were prepared and were photographed digitally to obtain the baseline L*, a*, b* values. Each sample was immersed in test solutions for 12 h which was equivalent time to 1 year of mouthrinse use, and the post-treatment images of the test materials were acquired. All L*, a*, b* values were analysed by a graphic software, and the total colour change (DeltaE*) of each specimen was calculated. Also the same colour analyses were performed on all test solutions to establish their colour parameters. Analysis of variance and Tukey's tests were used for statistical analyses and alpha was 0.05. All test solutions produced perceptible staining on the provisional material, with DeltaE values over 3.7. In both shades, hybrid rinse caused the highest staining (DeltaE=5.705), and was followed by chlorhexidine gluconate rinse, with DeltaE value of 4.120. The third highest staining was observed with benzydamine hydrochloride rinse (DeltaE=3.959), whereas the control caused the least staining (DeltaE=3.095). The lighter shade provisional material resulted with clinically observable staining even when immersed in distilled water; however, the dark shades showed clinically perceptible staining solely with the hybrid mouthrinse. In this study, the shade of the acrylic material was the determinator of the staining process. PMID:17371568

Cal, E; Güneri, P; Kose, T



Sperm viability staining in ecology and evolution: potential pitfalls  

Microsoft Academic Search

The causes and consequences of variation in sperm quality, survival and ageing are active areas of research in ecology and\\u000a evolution. In order to address these topics, many recent studies have measured sperm viability using fluorescent staining.\\u000a Although sperm viability staining has produced a number of interesting results, it has some potential pitfalls that have rarely\\u000a been discussed. In the

Luke Holman



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

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

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


Removing foxing stains from old paper at 157 nm  

Microsoft Academic Search

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

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



Complex III staining in blue native polyacrylamide gels  

Microsoft Academic Search

For more than a decade now blue native polyacrylamide gel electrophoresis (BN-PAGE) has been used for the study of the oxidative\\u000a phosphorylation (OXPHOS) complexes. Catalytic activities of complexes I, II, IV and V can be assessed, after separation by\\u000a gel electroforesis, by incubation of the BN-PAGE gel in specific staining solutions. However, until now, a reliable staining\\u000a method for testing

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



Gram Staining for the Treatment of Peritonsillar Abscess  

PubMed Central

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

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



Serum PABA and fluorescein in the course of Bz-Ty-PABA and pancreolauryl test as an index of exocrine pancreatic insufficiency  

Microsoft Academic Search

Forty-six subjects (20 chronic pancreatitis, 7 chronic liver disease, 7 recovered from acute pancreatitis, 2 Crohn's disease, and 10 healthy controls) classified by S-C test as having normal pancreatic function (26 subjects), or moderate (10 subjects) and severe (10 cases) pancreatic insufficiency, were given, on different days, 1 g of oral PABA or 348 mg of oral fluorescein dilaurate. At

G. Cavallini; W. Piubello; G. Brocco; R. Micciolo; G. Chech; G. Angelini; L. Benini; A. Riela; L. Dalle Molle; I. Vantini; L. A. Scuro



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

NASA Astrophysics Data System (ADS)

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

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



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

Microsoft Academic Search

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

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



Resolution of rabbit polyclonal anti-fluorescein Fab (IgG) fragments into subpopulations differing in affinity and spectral properties of bound ligand.  


Fab fragments derived from ten different IgG populations of hyperimmune rabbit polyclonal anti-fluorescein antibodies were further resolved into subfractions based on differences in time-dependent dissociation from an FITC-adsorbent in the presence of 0.1 M fluorescein at 4 degrees C. Fab fragments separated into subpopulations based on specific dissociation times of 0.1 day, 1.0 day, 10 days and 100 days from the adsorbent. Finally, after the 100 days elution step incubation with 6.0 M guanidine-HCl was included to determine total protein concentration of specific anti-fluorescein Fab fragments. Yields of specifically eluted Fab fragments ranged from 12.7 to 84.1% of the total Fab population originally incubated with the adsorbent. All Fab polyclonal populations and subpopulations analyzed quenched the fluorescence of the bound ligand by 90% or greater. None of the plots of protein concentration versus percent yield of the total specific antibody obtained for each of the five resolved fractions constituting a specific polyclonal population conformed to Gaussian distributions. All resolved Fab subpopulations retained bound fluorescein ligand that exhibited significant bathochromic shifts in absorbancy. Based on the extent of the red-shift the antibodies segregated into one of two general spectral families showing either a peak shift to 505-507 nm or to 518-520 nm. The red-shift to 518-520 nm appeared unique to rabbit anti-fluorescein antibodies, since corresponding large shifts have not been observed with antibodies derived from other species (e.g. mouse, rat, chicken, etc.). K(d) values determined for the resolved fractions confirmed a continuous progression in affinity from the 0.1day through the 100 days elution. Preliminary isoelectric focusing analyses revealed progressive selection for relatively more homogeneous fractions, especially in the 100 days resolved fraction. PMID:11483208

Voss, E W; Croney, J C; Jameson, D M



Crystalline hydrates and polymeric Ag(I) complexes of isomeric flexible double betaines 1,4-bis( n-picolyloxyl)benzene- N, N?-diacetate ( n = 2,3,4)  

Microsoft Academic Search

The isomeric flexible double betaines 1,4-bis(2-picolyloxyl)benzene-N,N?-diacetate (L1), 1,4-bis(3-picolyloxyl)benzene-N,N?-diacetate (L2) and 1,4-bis(4-picolyloxyl)benzene-N,N?-diacetate (L3) were used to react with AgNO3, affording coordination polymeric complexes {[Ag2L1](NO3)2·3.5H2O}n (1), {[Ag2L1](NO3)2·6H2O}n (2) and {[Ag3L3](NO3)3·3H2O}n (3). The crystal structures of the betaine hydrates L1·8H2O, L2·6H2O, L3·5H2O and the silver(I) complexes have been determined. L1·8H2O consists of a hydrogen-bonded three-dimensional network, whereas both L2·6H2O and L3·5H2O feature hydrogen

Lin-Ping Zhang; Chi-Keung Lam; Hai-Bin Song; Thomas C. W. Mak



Rapid Methods of Staining Bacterial Spores at Room Temperature  

PubMed Central

Lechtman, M. D. (University of Southern California, Los Angeles), J. W. Bartholomew, A. Phillips, and M. Russo. Rapid methods of staining bacterial spores at room temperature. J. Bacteriol. 89:848–854. 1965.—Spores of Bacillus subtilis var. niger were stained in 2 min at room temperature, after suitable pretreatment, with a dye reagent composed of 2% crystal violet in 1% phenol and 26% ethanol. Pretreatments included heat fixation to 260 C, mechanical rupture, and hydrolysis at room temperature in 44 n H3PO4 for 5 min, 33.4 n H3PO4 for 10 min, 12 n HCl for 5 sec, 6 n HCl for 2 min, 12 n HNO3 for 5 sec, and 6 n HNO3 for 60 sec. Acid hydrolysis at 60 C enabled the lowering of both acid concentration and time: 33.4 n H3PO4 for 15 sec, 25.9 n H3PO4 for 60 sec, 2 n HCl for 30 sec, 1 n HCl for 30 sec, 2 n HNO3 for 15 sec, and 1 n HNO3 for 30 sec. After acid treatment, 1 n NaOH was used as a neutralization agent. The cytological manifestations of these pretreatments, examined in an electron microscope after replication, showed definite degradation of spore coats, which probably explains the increase in dye permeability. The pretreatments were evaluated for use in a differential staining procedure for spores and vegetative cells. They were found to be too drastic in that they resulted in replacement of the primary dye by the 0.25% safranine counter stain in both vegetative cells and endospores. Less drastic pretreatments, such as 6 n HNO3 for 10 sec at room temperature, gave good differential stains, but failed to stain some free spores. The staining techniques above were evaluated with six species of Bacillus and were found to apply to all. Images

Lechtman, M. D.; Bartholomew, J. W.; Phillips, A.; Russo, M.



Demonstration of urinary eosinophils in Schistosoma haematobium: a comparative study among three different stains.  


Three stains, Hansel's stain, alkaline erythrocin B (AEB) and naphthalene black (NB), were used to demonstrate eosinophils in the urine of patients infected with Schistosoma haematobium. Hansel's stain was superior to the other two stains; it stained eosinophils bright red and their nuclei faint blue, and they were easily differentiated from neutrophils, lymphocytes, macrophages and epithelial cells. The method using AEB took longer than Hansel's stain and 10% of the specimens were lost during staining with this method. Like eosinophils, the neutrophils took up NB stain and their nuclei stained poorly with the counterstain. PMID:7687881

Eltoum, I A; Suliaman, S M; Ismail, B M; Ali, M M; Homeida, M M



Coomassie blue staining for high sensitivity gel-based proteomics.  


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. This article is part of a Special Issue entitled: From Genome to Proteome: Open Innovations. PMID:23428344

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



Mesothelin and GPR30 Staining Among a Spectrum of Pancreatic Epithelial Neoplasms  

Microsoft Academic Search

Introduction: Our study attempts to characterize mesothelin and GPR30 \\/ estrogen receptor (ER) staining in pancreatic pathology. Materials and Methods: Immunohistochemical staining for mesothelin, GPR30, and ER was performed on a variety of pancreatic lesions. Results: 24 of 42 (57%) adenocarcinomas stained for mesothelin, while 0 of 16 non-carcinomas (0%) stained (p = 0.0000784). 35 of 39 (90%) adenocarcinomas stained

Joseph P. Glass; Gulshan Parasher; Hugo Arias-Pulido; Rachel Donohue; Lisa A. Cerilli; Eric R. Prossnitz



Abelardo Gallego (1879–1930) and his contributions to histotechnology: The Gallego stains  

Microsoft Academic Search

The Gallego general tissue stain is used in histology and histopathology and stains beautifully connective tissue, in which all nuclei are stained in magenta red, epithelial cytoplasm in red–yellow, connective tissue is stained in brilliant green, muscle in olive green, and keratinized epithelium and blood in grass green. There is also Gallego's method for differential staining of elastic tissue that

Carlos Ortiz-Hidalgo



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


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

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



Determination of Trace Deoxyribonucleic Acid by Using Fluorescein Isothiocyanate-Phenosafranine as a Double-Luminescent Phosphorescence Probe  

PubMed Central

Using Pb2+ as ion perturber, phenosafranine (PF) and fluorescein isothiocyanate (FITC) could emit strong and stable room temperature phosphorescence (RTP) signal on the filter paper, respectively. When they were mixed, the phenomenon that the RTP signal of PF and FITC enhanced significantly was found. And 1.12 ag DNA spot?1 (sample volume was 0.40 ?L, corresponding concentration was 2.8?×?10–15 g mL–1) could cause the RTP signal of both PF and FITC to enhance sharply. The content of DNA was proportional to the ?Ip of PF and FITC in the system at 634 and 659 nm. Thus, a new solid substrate room temperature phosphorimetry (SSRTP) for the determination of trace DNA was established by using FITC-PF as double-luminescent phosphorescence probe. The detection limit (LD) of this method calculated by 3Sb/k was 14 zg DNA spot–1 for PF and 18 zg DNA spot–1 for FITC, respectively, showing high sensitivity. It has been applied to the determination of trace DNA in practical samples and the analysis results were in accordance with those of fluorescence probe. The reaction mechanism of SSRTP for the determination of trace DNA was also discussed.

Huang, Xiao-Mei; Liu, Zhen-Bo; Li, Fei-Ming; Lin, Li-Ping; Wang, Xin-Xing; Lin, Chang-Qing; Huang, Ya-Hong; Li, Zhi-Ming; Lin, Shao-Qin



N-hydroxysuccinimidyl fluorescein-O-acetate as a fluorescent derivatizing reagent for catecholamines in liquid chromatography.  


A new amine-reactive derivatizing reagent, N-hydroxysuccinimidyl fluorescein-O-acetate (SIFA), was developed for catecholamine (CA) analysis in liquid chromatography. The reactivity of this reagent with the CAs norepinephrine (NE), epinephrine (E), and dopamine (DA) was investigated in detail. In aqueous methanol containing 32 mmol/L pH 9.0 H3BO3-Na2B4O7 buffer, SIFA reacted with NE, E, and DA under mild conditions. The derivatives were separated in 20 min on a C18 column with a mobile phase of methanol/water (38:62, v/v) containing 10 mmol/L pH 5.0 H3cit-Na2HPO4 buffer. At lambda(ex)/lambda(em) = 490/516 nm, the detection limits were 3.2, 12, and 56 fmol, respectively, with a signal-to-noise ratio of 3, which were comparable to those using 1,2-diphenylethylenediamine as the derivatizing reagent for CA analysis. Amino acids, aliphatic amines, and alcohols had no obvious interference with the determination. The proposed method has been applied to the determination of CAs in human urine, with recoveries of 95.3-103.9%. PMID:10847605

Wang, H; Li, J; Liu, X; Yang, T X; Zhang, H S



Grading of Age-Related Macular Degeneration: Comparison between Color Fundus Photography, Fluorescein Angiography, and Spectral Domain Optical Coherence Tomography  

PubMed Central

Purpose. To compare color fundus photography (FP), fluorescein angiography (FA), and spectral domain optical coherence tomography (SDOCT) for the detection of age-related macular degeneration (AMD), choroidal neovascularisation (CNV), and CNV activity. Methods. FPs, FAs, and SDOCT volume scans from 120 eyes of 66 AMD and control patients were randomly collected. Control eyes were required to show no AMD, but other retinal pathology was allowed. The presence of drusen, pigmentary changes, CNV, and signs for CNV activity was independently analyzed for all imaging modalities. Results. AMD was diagnosed based on FP in 75 eyes. SDOCT and FA showed sensitivity (specificity) of 89% (76%) and 92% (82%), respectively. CNV was present on FA in 68 eyes. Sensitivity (specificity) was 78% (100%) for FP and 94% (98%) for SDOCT. CNV activity was detected by SDOCT or FA in 60 eyes with an agreement in 46 eyes. Sensitivity was 88% for SDOCT and 88% for FA. FP showed sensitivity of 38% and specificity of 98%. Conclusions. CNV lesions and activity may be missed by FP alone, but FP may help identifying drusen and pigmentary changes. SDOCT is highly sensitive for the detection of AMD, CNV, and CNV activity; however, it cannot fully replace FA.

Mokwa, Nils F.; Keane, Pearse A.; Kirchhof, Bernd; Sadda, Srinivas R.



Stain length passive dosimeter for monitoring of carbon monoxide  

SciTech Connect

A new passive dosimeter for the personal monitoring of CO exposure in the workplace has been developed. This new type of sampling device does not require follow-up analysis of the collected sample to determine the exposure. Rather, the time-weighted average concentration is determined directly from the length of a coloured stain which is produced instantaneously during the exposure period in a specially prepared indicator tube. The stain length is a function of both contaminant concentration and exposure time. The effects of CO concentration in the range 0.5-2.0 x TLV on the response of the dosimeter were evaluated in a dynamic exposure chamber. Data were fitted to the appropriate model equation with a correlation coefficient of 0.968. By eliminating the need for follow-up analysis, this stain-length dosimeter significantly reduces the cost of monitoring.

Gonzalez, L.A.; Sefton, M.V.



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


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

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



Evaluation of peroxidase activity by alpha-naphthol/pyronine staining compared with benzidine staining in 101 acute leukemia cases.  


Cytochemical detection of myeloperoxidase (MPO) activity, a strong marker for myeloid differentiation, is usually performed by benzidine dihydrochloride staining, with the threshold at 3%. Several reports have demonstrated the potential toxicity of benzidine, and bans have been issued, under French law, prohibiting female technicians from being exposed to the aromatic hydrocarbon group, including benzidine. The aim of this study was to test an alpha-naphthol and pyronine-based substitute using a standardized kit (MYELOPEROXIDASE KIT, RAL [Réactifs RAL, Martillac, France]) to measure MPO activity in blast cells. This prospective, multicenter study made it possible to analyze 101 acute leukemia (AL) cases; it has also demonstrated both the 96% specificity and the 99% sensitivity of the method, with a threshold for positive staining of 3%, as well as good correlation (r = 0.95) between the staining method tested and the benzidine staining method. When using the alpha-naphthol/pyronine-based staining for MPO, the mean number of positive blast cells is statistically lower than that obtained using benzidine, but without incidence on AL classification. These results allow us to conclude that this method makes it possible to classify acute blood diseases by measuring MPO activity using reagents permitted by law, according to a standardized and reproducible protocol. PMID:21097443

Latger-Cannard, Véronique; Bardet, Valérie; Malet, Michèle; Lagrange, Monique; Empereur, Fabienne; Fenneteau, Odile



Staining methods applied to glycol methacrylate embedded tissue sections.  


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

Cerri, P S; Sasso-Cerri, E



Modeling of alkane emissions from a wood stain  

SciTech Connect

The article discusses full-scale residential house tests to evaluate the effects of organic emissions from a wood finishing product--wood stain--on indoor air quality (IAQ). The test house concentrations of three alkane species, nonane, decane, and undecane, were measured as a function of time after the application of the wood stain. It was found that the test house concentrations can be simulated by an integrated IAQ model which takes into consideration source, sink, and ventilation effects. The alkane emissions were controlled by an evaporation-like process.

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



Toward digital staining using imaging mass spectrometry and random forests.  


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

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



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



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


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. PMID:23666356

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



Study of Bacillus Subtilis Endospores in Soil by Use of a Modified Endospore Stain.  

National Technical Information Service (NTIS)

The Schaeffer-Fulton endospore stain was modified so that it would stain Bacillus subtilis endospores in soil smears. The modified stain differentiated among dormant spores, spores undergoing activation, and spores which had germinated but had not yet sho...

D. A. Mormak L. E. Casida



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

SciTech Connect

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

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



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 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

Tom C. Pearson




EPA Science Inventory

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



EPA Science Inventory

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


Immunohistochemically Stained Activated Eosinophils in Sputum in Patients with Asthma  

Microsoft Academic Search

Background: Eosinophils play an important role in asthmatic airway inflammation. Monoclonal antibody EG2 has been considered to identify activated eosinophils. Objective: The present study was aimed to investigate whether immunohistochemically stained EG2+ eosinophils in sputum reflect the severity of asthma. Methods: Sputum was obtained in 23 asthmatic patients, of whom 13 patients were examined before and after antiasthma treatment including

An-Soo Jang; Inseon-S Choi; Chang-Soo Park



LacCur stain for detection of mucin in adenocarcinoma.  


Identification of cytoplasmic mucin, usually by Mayer's mucicarmine stain, is one of the criteria to diagnose adenocarcinoma. The inexpensive LacCur stain, made up of Curcuma longa (khamin-shan) and secreta of Laccifer lacca (krang) has been introduced. The aim of this study was to compare the Mayer's mucicarmine and LacCur stains in the detection of mucin material. The specimens included 17 adenocarcinomas of the stomach, 16 of the colon, 18 of the lung, 16 of the breast, and 12 of the bile duct. Squamous cell carcinoma and hepatocellular carcinoma (altogether 20 cases) were set as negative control. Like Mayer's mucicarmine, LacCur was capable of detecting of intracytoplasmic mucin in all adenocarcinomas of the stomach, colon and bile duct, and revealed mucin substance in 15/18 and 11/16 cases of specimens from the lung and breast, respectively. The negative control group showed a negative result. Although a little more time required in preparation, the LacCur stain is simple and very economical. PMID:14971540

Wattanasirmkit, Vanee; Tanboon, Jantima; Shuangshoti, Somruetai; Shuangshoti, Shanop



Analysis of Oxiclean: An Interesting Comparison of Percarbonate Stain Removers  

ERIC Educational Resources Information Center

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

Bracken, Jeffrey D.; Tietz, David



Histochemistry of the Gram-staining Reaction for Microorganisms  

Microsoft Academic Search

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

H. Henry; M. Stacey



Feasibility of digitally stained multimodal confocal mosaics to simulate histopathology  

Microsoft Academic Search

Fluorescence confocal mosaicing microscopy of tissue biopsies stained with acridine orange has been shown to accurately identify tumors and with an overall sensitivity of 96.6% and specificity of 89.2%. However, fluorescence shows only nuclear detail similar to hematoxylin in histopathology and does not show collagen or cytoplasm, which may provide necessary negative contrast information similar to eosin used in histopathology.

Daniel S. Gareau



Incidence of homogeneously staining regions in non-Hodgkin lymphomas  

Microsoft Academic Search

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

E. Arranz; M. Robledo; B. Martínez; J. Gallego; A. Román; C. Rivas; J. Benítez



The involvement of nucleosomes in Giemsa staining of chromosomes  

Microsoft Academic Search

Summary A new hypothesis is proposed on the involvement of nucleosomes in Giemsa banding of chromosomes. Giemsa staining as well as the concomitant swelling can be explained as an insertion of the triple charged hydrophobic dye complex between the negatively-charged supercoiled helical DNA and the denatured histone cores of the nucleosomes still present in the fixed chromosomes. New cytochemical data

P. van Duijn; A. C. Prooijen-Knegt; M. Ploeg



Very bright europium complexes that stain cellular mitochondria.  


The synthesis, structure and photophysical properties of a series of highly emissive europium complexes is reported. Certain complexes enter mammalian cells by macropinocytosis and stain the mitochondria selectively, allowing observation of the Eu emission in cellulo by time-gated spectral imaging. PMID:23336102

Walton, James W; Bourdolle, Adrien; Butler, Stephen J; Soulie, Marine; Delbianco, Martina; McMahon, Brian K; Pal, Robert; Puschmann, Horst; Zwier, Jurriaan M; Lamarque, Laurent; Maury, Olivier; Andraud, Chantal; Parker, David



A New Method in Staining Bacteria on Microscope Slides  

Microsoft Academic Search

A NEW method of providing dyes for some of the commoner staining procedures has been developed and tested. A small volume of the required dye solution is prepared according to the appropriate formula. The fluid is placed in a large dish, and rounds of No. 4 Whatman filter paper are soaked in it. After a few seconds the paper is

P. H. H. Gray



Histochemical staining of orbicularis oculi muscle in ectropion and entropion  

Microsoft Academic Search

A histochemical study of orbicularis oculi was undertaken to test the hypothesis that there is a difference in the percentage and size of muscle fibre types which accounts for the development of involutional ectropion or entropion. Wedge excisions from lower lids of patients undergoing repair of these conditions were frozen-sectioned and stained histochemically to reveal muscle fibre types. Five ectropion

Ruth M Manners; Roy O Weller



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



Discrimination of homologous chromosomes of maize with giemsa staining  

Microsoft Academic Search

A Giemsa method is described for the identification of somatic chromosomes of maize (Zea mays L.). Three inbred stocks and their hybrids were examined. The Giemsa method proved to be useful for the characterisation of different stocks. On the basis of the different staining properties of homologous chromosomes, the parental and grandparental chromosomes were identified in the hybrids.

Gy Hadlaczky; L Kálmán



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



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



Image analysis of dye stained patterns in soils  

NASA Astrophysics Data System (ADS)

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

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



Surgery for the extensive facial port-wine stain?  

Microsoft Academic Search

In contrast to true hemangiomas, which are vascular neoplasms, port-wine stains (PWS) are vascular malformations. They consist of mature teleangiectatic vessels in the dermis and the adjacent subcutis. The series of 50 patients here reported have been treated by subtotal excision of their PWS, covering the resulting defects with carefully selected full-thickness skin grafts.

L. Clodius



Non-staining, non-sticking styrenic polymers  

SciTech Connect

A method is described for preparing a non-staining, non-sticking, high impact polystyrene, which comprises: (a) a solution formed by dissolving a rubber in a mixture consisting essentially of a styrene, a polysiloxane and a solvent; (b) heating the resultant solution to graft polymerize the styrene onto the rubber; and (c) separating the solvent and any unreacted styrene present.

Gunesin, B.Z.



Observation of Soft Contact Lens Disinfection with Fluorescent Metabolic Stains  

PubMed Central

A rapid fluorescent staining method using a tetrazolium dye and propidium iodide for the in-use assessment of disinfection of Pseudomonas aeruginosa biofilms on soft contact lenses showed that 11 to 13% of cells on lenses remained actively respiring and recoverable by culture methods after 30 min of exposure to 3% hydrogen peroxide.

Gavin, J.; Button, N. F.; Watson-Craik, I. A.; Logan, N. A.



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

PubMed Central

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

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



Urine fluorescence using a Wood's lamp to detect the antifreeze additive sodium fluorescein: A qualitative adjunctive test in suspected ethylene glycol ingestions  

Microsoft Academic Search

Antifreeze ingestions require rapid and accurate differential diagnosis to prevent fatal outcomes. Sodium fluorescein is added to some commercial antifreeze preparations (ethylene glycol) to a final concentration of approximately 20 micrograms\\/mL as a colorant to aid in detection of automobile cooling-system leaks. For an adult human being, a potentially toxic volume of antifreeze is 30 mL, which contains 0.4 to

M. L. Winter; M. D. Ellis; W. R. Snodgrass



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

NASA Astrophysics Data System (ADS)

The silica-encapsulated luminescent lanthanide nanoparticles have been developed for the selective tagging of a wide range of important targets in recent years, however, they are mainly limited to europium and terbium compounds. In this work, two types of europium-containing dual-luminophore silica nanoparticles, silica-encapsulated CdTe quantum dots (CdTe QDs)-BHHCT-Eu3 + complex nanoparticles and BHHCT-Eu3 + surface-bound silica-encapsulated fluorescein isothiocyanate (FITC) nanoparticles (BHHCT: 4, 4'-bis(1'', 1'', 1'', 2'', 2'', 3'', 3''-heptafluoro-4'', 6''-hexanedion-6''-yl)chlorosulfo-o-terphenyl), were successfully prepared using a water-in-oil (W/O) reverse microemulsion method. The results of transmission electron microscopy and luminescence spectroscopy characterizations indicate that the two types of nanoparticles are all monodisperse, spherical and uniform in size (~50 nm in diameter), and have well-resolved and stable dual luminescence emission properties. The CdTe QDs-BHHCT-Eu3 + nanoparticles can be excited at 365 nm to give dual-emission peaks at 535 and 610 nm, and the FITC-BHHCT-Eu3 + nanoparticles can be excited at 335 nm to give dual-emission peaks at 515 and 610 nm. The luminescence response investigations of the nanoparticles to different metal ions indicate that the new nanoparticles can be used as ratiometric luminescent sensing probes for the selective detection of Cu2 + and Fe2 + ions, respectively. The performance of the nanoparticle probe for metal ion detection was investigated.

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



Percutaneous penetration of fluorescein isothiocyanate-dextrans and the mechanism for enhancement effect of enhancers on the intercellular penetration.  


To identify the mechanism involved in the enhancement effect of enhancers on the intercellular penetration of large polar molecules, the skin penetration of fluorescein isothiocyanate (FITC)-dextrans (average molecular weight; 4400, 9400, and 69000 Da) and the lipid removal from the intercellular spaces by enhancers were studied using hairless rat skin. Pretreatment of hairless rat skin with enhancers such as n-octanol (20%), laurocapram (2%), isopropylmyristate (IPM, 20%), oleic acid (5%) and cineol (2%), which are water-immiscible, significantly enhanced the flux of FITC-dextrans, while pretreatment with water-miscible enhancers, i.e. dimethyl sulfoxide (DMSO, 5%) and N-methyl-2-pyrrolidone (NMP) did not increase the flux compared with the control. The penetration of FITC-dextrans was approximately size dependent. n-Octanol, laurocapram, IPM and oleic acid dramatically removed ceramides which are the intercellular lipids, whereas NMP and DMSO partly extracted the sphingolipids. A linear relationship was observed between the flux and removal of ceramides (p < 0.01), indicating that the removal of intercellular lipids would cause dramatic dilations between adherent cornified cells and enhance the penetration through the intercellular pathways. When the penetration of FITC-dextrans through Wistar rat skin was compared with that via hairless rat skin, the steady state flux of FITC-dextrans through Wistar rat skin pretreated with water-immiscible enhancers was 1.2- to 4.9-fold higher, suggesting that the penetration of large polar molecules through follicles may play at least some role in the percutaneous absorption. PMID:8593481

Ogiso, T; Paku, T; Iwaki, M; Tanino, T



An evaluation of Com-B27, a fluorescein-conjugated mouse anti-HLA-B27 reagent.  


We evaluated the HLA-B27 typing reagent Com-B27, which is claimed to show minimal cross-reactivity with HLA-B7, for use in flow cytometry-based typing. This combination reagent consists of the fluorescein-conjugated HLA-B27 mouse monoclonal antibody ABC-m3 and a non-conjugated HLA-B7 monoclonal antibody that is claimed to block the reactivity of ABC-m3 to B7 without affecting its reactivity to B27. It reacted well with B27 (B*2702, B*2705) and B2708 [mean median channel fluorescence intensity (MCFI) 8.40] and weakly with B7 (including cells from homozygous B*07 donors), B42/B73 and B22/B37/B44 reference cells (mean MCFI 2.05, 3.09 and 1.10, respectively). It showed a uniform discrimination between B7 and B27/B2708 with no 'overlap' in MCFI values, which was seen with the standard ABC-m3 antibody. There was complete agreement when our standard three-antibody-based B27/B2708 flow cytometry assay and the Com-B27 reagent alone were used to independently assign B27/B2708 status to 651 random patients. Thus, the Com-B27 reagent provided improved discrimination between B7 and B27/B2708 over the ABC-m3 antibody, and its B27/B2708 assignments were comparable with our standard flow cytometry assay. However, for consistently reliable B27/B2708 typing we continue to recommend the use of a minimum of two B27 reagents in a protocol that includes DNA-based testing of 'equivocal' B27/B2708 assignments. PMID:12919288

Coates, E; Rees, T J; Darke, C



Association of Fluorescein Angiographic Features with Visual Acuity and with Optical Coherence Tomographic and Stereoscopic Color Fundus Photographic Features of Diabetic Macular Edema in a Randomized Clinical Trial  

PubMed Central

Background Fluorescein angiography (FA) has been performed as part of the management of diabetic macular edema (DME) for many years. Its current role relative to the role of optical coherence tomography (OCT) is not well defined. Purpose To evaluate the associations of FA features with visual acuity, and with OCT, and fundus photographic characteristics in eyes with DME. Methods In a clinical trial, conducted by the Diabetic Retinopathy Clinical Research Network to compare two methods of laser photocoagulation to treat DME, FA (film and digital), color photographs, OCT, and visual acuity measurements were obtained at baseline and at 1 year. Grading of morphologic features was performed at a reading center. Reproducibility of FAs was assessed and the correlations of FA features with visual acuity, OCT, and color photograph features were computed. Results From 79 clinical sites, data of 323 study eyes and 203 fellow non-study eyes were analyzed. Fluorescein leakage area at baseline was associated with reduced visual acuity, increased OCT measures of retinal thickness and volume, and color photographic measurements of retinal thickening (r = 0.33 – 0.58). No important associations were found with changes from baseline to 12 months in these parameters or with any of the other variables analyzed. Conclusions Fluorescein leakage is associated with visual acuity and some OCT and color photographic variables. We did not identify any unique FA variables that had a stronger association with visual acuity than OCT measures of retinal thickness. These data may be useful to investigators planning future DME clinical trials.

Danis, Ronald P.; Scott, Ingrid U.; Qin, Haijing; Altaweel, Michael M.; Bressler, Neil M.; Bressler, Susan B.; Browning, David J.; Kollman, Craig



[Bertalanffy-like fluorescence staining with 3-dimethylamino-6-methoxyacridine].  


Three new acridine dyes, 3-dimethylamino-6-methoxyacridine 1, 3-amino-6-methoxyacridine 2 and 3-amino-7-methoxyacridine 3, have been prepared and tested as fluorochromes of LM- and HeLa-cells. The dyes are basic compounds (pKA: 1 8,76; 2 8,01; 3 7,65) and form cations in neutral or acidic aqueous solutions by addition of a proton to the aza-nitrogen atom of the heterocycle. The fluorochromes stain fixed LM- and HeLa-cells at pH = 6. The fluorescence shows metachromasy similar to the staining with acridine orange AO according to the technique of Bertalanffy. But there is less fading of the fluorescence. The dye 1 is the most suitable fluorochrome of the series. It was studied in detail. Using optimized staining conditions the fluorescence of the nucleus is yellow-green that of the cytoplasm and the nucleoli orange or brownish-red. Enzymatic digestion experiments show that the dye cations are bound to DNA in the nucleus and to RNA in the cytoplasm or nucleoli. The absorption and emission spectra of the stained cells have been studied by means of microspectrophotometry. The absorption spectra of the nucleus and the cytoplasm are very similar. The maximum of the long wave length absorption of both occurs at 21400 cm-1 (467 nm) with a shoulder at ca 20100 cm-1 (498 nm). The fluorescence spectra of nucleus and cytoplasm of metachromatically stained cells are different. The emission maximum of the cytoplasm and nucleoli, 16200 cm-1 (617 nm), is red-shifted relative to the maximum of the nucleus, 18200 cm-1 (549 nm). This shift causes the metachromatic fluorescence effect. In addition we studied the concentration dependence of the absorption and fluorescence spectra of the cation 1 in aqueous solution, pH = 6, in the concentration range 6 X 10(-6)-6 X 10(-4) M. Shape and maximum of the long wave length absorption and emission depend only slightly on the concentration: Mean value of absorption maximum ca 21500 cm-1 (465 nm), shoulder at ca 20300 cm-1 (493 nm), fluorescence maximum ca 18300 cm-1 (547 nm). With growing concentration diminishes the molar absorptivity. This decrease in absorptivity and isosbestic points in the absorption spectra indicate the formation of dimers with growing dye concentration. The absorption spectra of the metachromatically stained cells and of the dye in aqueous solution are very similar.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:6203878

Petschel, K; Naujok, A; Kempter, P; Seiffert, W; Zimmermann, H W



Comparative Analysis of an Improved Thioflavin-S Stain, Gallyas Silver Stain, and Immunohistochemistry for Neurofibrillary Tangle Demonstration on the Same Sections  

Microsoft Academic Search

SUMMARY An improved thioflavin-S stain, Gallyas silver stain, and two immunostainings were quantitatively compared for demonstration of neurofibrillary tangles (NFTs) on the same sections. Sections of hippocampal formation from seven cases of Alzheimer's disease (AD) were immunofluorescently stained with a commercially available polyclonal NFT anti- body or a PHF-1 monoclonal antibody, followed by an improved thioflavin-S stain, and fi- nally

Anyang Sun; Xuan V. Nguyen; Guoying Bing



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


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

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



Imaging port wine stains by fiber optical coherence tomography  

NASA Astrophysics Data System (ADS)

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

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



[NBI magnifying endoscopic classification using crystal violet staining].  


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

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



Solving the mystery of the Colorado Brown Stain.  


The life and work of Dr. Frederick S. McKay in solving the mystery of the Colorado Brown Stain changed the objectives of restorative and preventive dentistry. McKay was an intellectually diversified man whose personal interests ranged from economics to opera. Professionally his strong commitment to research led to dedicate thirty years of his life to the search for the mysterious agent that caused the Colorado Brown Stain which mottled but also produced caries-free teeth. His discovery of fluoride in drinking water and its effect on enamel was a critical breakthrough in understanding the etiology and prevention of dental caries. This discovery is the foundation for water fluoridation which is the single most effective public health measure to inhibit tooth decay. PMID:9468893

Peterson, J



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.



Development of Cell Staining Technique for X-Ray Microscopy  

SciTech Connect

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

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



Identification of active fluorescence stained bacteria by Raman spectroscopy  

NASA Astrophysics Data System (ADS)

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

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



Simple differential Giemsa staining of sister chromatids after treatment with photosensitive dyes and exposure to light and the mechanism of staining  

Microsoft Academic Search

The essential steps of the 33258 Hoechst-Giemsa method for differential chromatid staining consist of (1) 33258 Hoechst treatment, (2) exposure to light, and (3) Giemsa staining. The staining was shown to be a function of the concentration of 33258 Hoechst and the light exposure. The dye was successfully replaced by various metachromatic dyes such as thionine. Two simple methods are

Keiko Goto; T. Akematsu; H. Shimazu; T. Sugiyama



Tunable filter-based multispectral imaging for detection of blood stains on construction material substrates. Part 1. Developing blood stain discrimination criteria.  


In this article, we establish blood stain detection criteria that are less substrate dependent for use in a liquid crystal tunable filter-based multispectral-imaging system. Kubelka-Munk (KM) theory is applied to transform the acquired stains' reflectance spectra into the less substrate dependent spectra. Chosen spectral parameters are extracted from the KM absorbance spectra of several stain samples on several substrates. Blood discrimination criteria based upon those spectral parameters are then established from empirical data, tested, and refined. In our newly invented method, instead of introducing conventional contrast enhancement on the blood stain image, blood stain determination is executed mathematically via Boolean logic, resulting in more discriminative blood stain identification. This proposed approach allows for nondestructive, quick, discriminative, and easy-to-improve presumptive blood stain detection. Experimental results confirm that our blood stain discrimination criteria can be used to locate blood stains on several construction materials with high precision. True positive rates (sensitivity) from 0.60 to 0.95 are achieved depending on blood stain faintness and substrate types. Also, true negative rates (specificity) between 0.55 and 0.96 and identification time of 4-5 min are accomplished, respectively. The established blood stain discrimination criteria will be incorporated in a real blood stain detection system in part 2 of this article, where system design and considerations as well as speed issues are discussed. PMID:23052077

Janchaysang, Suwatwong; Sumriddetchkajorn, Sarun; Buranasiri, Prathan



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

Microsoft Academic Search

Blood stains can be crucial in reconstructing crime events. However, no reliable methods are currently available to establish the age of a blood stain on the crime scene. We show that determining the fractions of three hemoglobin derivatives in a blood stain at various ages enables relating these time varying fractions to the age of the blood stain. Application of

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



A Fluorescent Gram Stain for Flow Cytometry and Epifluorescence Microscopy  

Microsoft Academic Search

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




Quantitative sputum gram stains in chronic bronchial disease  

Microsoft Academic Search

The assessment of bacterial flora of the bronchial system can provide useful information for determining the presence of acute\\u000a bacterial infections in patients with chronic bronchial disease. The authors examined the value of quantitative sputum gram\\u000a stains performed in patients during acute bronchial exacerbations, recovery from such exacerbations, acute allergic exacerbations\\u000a of chronic extrinsic asthma, and a stable period. The

W. Baigelman; S. Chodosh; D. Pizzuto; T. Sadow



Polyphenol oxidase activity staining in polyacrylamide electrophoresis gels.  


An analytical method allowing the detection of polyphenol oxidase activity on polyacrylamide gel electrophoresis (PAGE) is described. The method is rapid, sensitive and specific and is based on a coupling reaction between 4-tert-butyl-o-benzoquinone and the aromatic amine, 4-amino-N,N-diethylaniline sulphate. Catecholase activity of polyphenol oxidase appears as blue stained bands on a colourless background. PMID:9178091

Rescigno, A; Sollai, F; Rinaldi, A C; Soddu, G; Sanjust, E



Permanence of Fungi-Fluor epifluorescence stain to detect microsporidia  

Microsoft Academic Search

OBJECTIVE:Epifluorescence microscopy, a methodology for the screening of bodily fluids and tissue specimens for microsporidia species, was directed to evaluate the retention of epifluorescence of fixed and stained specimens over time.METHODS:Thirty samples of stool, bodily fluids, duodenal touch preparations, and biopsies, were tested for the retention of their epifluoresence using the Fungi-Fluor procedure. Specimens were examined under a 330- to

O. G. W. Berlin; C. N. Conteas; L. R. Ash; F. Sorvillo; C. V. Jacob; J. Yatabe; J. B. Peter



CMA staining analysis of chromosomes in several species of Aurantioideae  

Microsoft Academic Search

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

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



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.




A combination of sister chromatid differential staining and giemsa banding  

Microsoft Academic Search

Summary  We report a procedure for combining sister chromatid differential staining and G banding in the same metaphase plate. Mammalian\\u000a cells in culture are grown in medium containing 5-bromodeoxyuridine for two cell cycles, and conventional air-dried preparations\\u000a are made. The slides are treated with a trypsin or a urea solution the same way as for regular G banding. This method is

S. Pathak; A. D. Stock; A. Lusby



Specific silver staining of experimentally undercondensed chromosome regions  

Microsoft Academic Search

Treatment of human and mouse cell cultures with DNA binding AT-specific compounds and with some base analogues induced distinct undercondensations in several heterochromatic chromosome regions. All those heterochromatic regions undercondensed by AT-specific DNA ligands (distamycin A, DAPI, Hoechst 33258) could be heavily labeled with the silver(Ag)-staining technique; but the heterochromatic regions undercondensed with the cytidine analogue 5-azacytidine were Ag-negative. In

T. Haaf; H. Weis; D. Schindler; M. Schmid



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



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


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

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



A new staining method of astrocytes for paraffin section.  


A new method of staining astrocytes in formation-fixed, paraffin-embedded sections was devised: (1) fix them in 5% mercuric chloride solution for 30 min to 1 h at 56 degrees C, (2) then place in 0.5% iodine alcohol for 5 min followed by placing in 0.5% sodium thiosulfate for 5 min, (3) immerse in 0.25% potassium permanganate for 3 min, (4) place in 2% oxalic acid for 2 min. (5) mordant in 2% iron alum for 45 s, and (6) place in 2% silver nitrate solution for 30 min. The next step is impregnation in ammoniacal silver solution for 10 -- 15 min at 56 degrees C, followed by reduction in neutral formalin and 2% iron alum, toning in 0.2% gold chloride, and fixing in 5% sodium thiosulfate. Pathological astrocytes of fibrillary and protoplasmic types were distincly demonstrated, although nerve cells and nuclei of oligodendrocytes and microglial cells were also faintly stained. Thus, the staining for paraffin sections is fairly selective for astrocytes in pathological states. PMID:6153491

Kitoh, T; Matsushita, M



Nile red: a selective fluorescent stain for intracellular lipid droplets  

PubMed Central

We report that the dye nile red, 9-diethylamino-5H- benzo[alpha]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. Better selectivity for cytoplasmic lipid droplets was obtained when the cells were viewed for yellow-gold fluorescence (excitation, 450-500 nm; emission, greater than 528 nm) rather than red fluorescence (excitation, 515-560 nm; emission, greater than 590 nm). Nile red-stained, lipid droplet-filled macrophages exhibited greater fluorescence intensity than did nile red- stained control macrophages, and the two cell populations could be differentiated and analyzed by flow cytofluorometry. Such analyses could be performed with either yellow-gold or red fluorescence, but when few lipid droplets per cell were present, the yellow-gold fluorescence was more discriminating. Nile red exhibits properties of a near-ideal lysochrome. It is strongly fluorescent, but only in the presence of a hydrophobic environment. The dye is very soluble in the lipids it is intended to show, and it does not interact with any tissue constituent except by solution. Nile red can be applied to cells in an aqueous medium, and it does not dissolve the lipids it is supposed to reveal.



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.

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



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

PubMed Central

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

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



Antibody-antigen binding constants determined in solution-phase with the threshold membrane-capture system: binding constants for anti-fluorescein, anti-saxitoxin, and anti-ricin antibodies.  


Affinities of various monoclonal and polyclonal antibodies for fluorescein-containing antigens, saxitoxin and ricin, were determined by using a light addressable potentiometric sensor-based system (Threshold). The dissociation constants, determined from Scatchard plots, ranged from 2 x 10(-7) to approximately 3 x 10(-12) M. Dissociation constants for fluorescein and saxitoxin were compared with values determined by independent means. This technique was found to be quick, simple, reproducible, and accurate. PMID:8203727

Dill, K; Lin, M; Poteras, C; Fraser, C; Hafeman, D G; Owicki, J C; Olson, J D



Evaluation of a Rapid Fluorescent Staining Method for Detection of Mycobacteria in Clinical Specimens ?  

PubMed Central

Rapid detection of mycobacterial disease is essential. Using multiple specimen types and concentrations of mycobacteria, we compared two commercial auramine O stains. The more rapid stain permitted consistent acid-fast bacillus quantitation and exhibited less debris staining, and the staining procedure required less time (?2 min) to perform. These results suggest that the rapid stain may be more cost-effective and efficient for use in clinical laboratories.

Hendry, Cindy; Dionne, Kim; Hedgepeth, Annie; Carroll, Karen; Parrish, Nicole



[Accidental staining of corneal nerves by methylene blue].  


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

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



Identification of homogeneously staining regions in leukemia patients  

PubMed Central

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

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



Epithelial membrane antigen staining patterns of histiocytic lesions.  


Epithelial membrane antigen (EMA) appears to be a marker of activation, proliferation, and/or neoplasia in some epithelial and nonepithelial cells, including histiocytes. We performed immunoperoxidase stains for EMA on a variety of histiocytic lesion specimens, including specimens from two cases of interdigitating reticulum cell lymphoma, 12 cases of histiocytosis X, seven cases of sarcoidosis, five cases of granuloma annulare, 13 juvenile xanthogranulomas, two reticulohistiocytomas, five xanthelasmas, three dermatopathic lymph nodes, and three foreign body reactions. Only the two cases of interdigitating reticulum cell lymphoma and two of the 12 cases of histiocytosis X exhibited significant EMA positivity. These findings may prove useful in the differential diagnosis of histiocytic lesions. PMID:3103582

Rabkin, M S; Kjeldsberg, C R



News from the biological stain commission no. 14.  


In the 14(th) issue of News from the Biological Stain Commission (BSC) the BSC's International Affairs Committee presents information from the meetings of ISO/TC 212/WG 3, In vitro diagnostic products, and from the final plenary meeting of ISO/TC 212, Clinical laboratory testing and in vitro diagnostic test systems, held on 17-19 October 2011 in Las Vegas, NV. Information is also presented from the 26th meeting of CEN/TC 140, In vitro diagnostic medical devices, held on 5 December 2011 in Berlin, Germany. PMID:23461718

Lyon, Hans O



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.



Computerized image analysis of iron-stained macrophages.  


Analysis of iron levels in single cells is critical to understand the consequences of impaired regulation of iron homeostasis. Here we establish a method to analyze intracellular iron deposits by computerized image analysis of Prussian blue-stained alveolar macrophages as a test system. We efficiently detected small differences in macrophage steady-state iron levels in Hfe (-/-) mice as well as inflammation-induced iron sequestration upon lipopolysaccharide instillation. In conclusion, computerized image analysis of single cells is a robust and reproducible tool suitable for iron measurements in small sample sets with limited cell yield. PMID:23592271

Benesova, Karolina; Schaefer, Sebastian M; Mall, Marcus A; Muckenthaler, Martina U



DNA extraction and amplification from Giemsa-stained blood smears.  


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

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



DAPI staining and fluorescence microscopy techniques for phytoplasmas.  


The 4',6-diamidino-2-phenylindole (DAPI) stain technique is a simple method that was developed for confirming the presence of phytoplasmas in hand-cut or freezing microtome sections of infected tissues. DAPI binds AT-rich DNA preferentially, so that phytoplasmas, localized among phloem cells, can be visualized in a fluorescence microscope. The procedure is quick, easy to use, inexpensive, and can be used as a preliminary or quantitative method to detect or quantify phytoplasma-like bodies in infected plants. PMID:22987410

Andrade, Nancy M; Arismendi, Nolberto L



Differentiation Between Presumed Ocular Histoplasmosis Syndrome and Multifocal Choroiditis With Panuveitis Based on Morphology of Photographed Fundus Lesions and Fluorescein Angiography  

Microsoft Academic Search

were available, observer A and observer B k values against the gold standard were 0.625 (95% CI, 0.270-0.980) and 0.588 (95% CI, 0.235-0.940), respectively. The pictures- only k values for observer A vs observer B were 0.582 (95% CI, 0.316-0.848) with indeterminate patients factored in and 1.0 (95% CI, 1.0-1.0) when indeterminate patients were excluded. Pictures and fluorescein angiogram k

Jeffrey R. Parnell; Lee M. Jampol; Lawrence A. Yannuzzi; J. Donald; M. Gass; Michael K. Tittl



Chiral separation of amino acids derivatised with fluorescein isothiocyanate by single isomer derivatives 3-monodeoxy-3-monoamino-?- and ?-cyclodextrins: the effect of the cavity size.  


Thirteen enantiomeric pairs of ?-amino acids derivatised with fluorescein isothiocyanate (FITC-AAs) were separated in capillary electrophoresis (CE) using as chiral selectors the single isomer derivatives (SIDs) 3-monodeoxy-3-monoamino-?- and ?-cyclodextrins. The chiral separation data obtained by these strictly homologous compounds, show different behaviours, allowing to hypothesise a possible structure of the obtained selector-analyte complexes, as well as highlighting the crucial role of the cavity size and the significant effects on the resolution obtained by small differences in the structural characteristics of these analytes. PMID:22939205

Giuffrida, Alessandro; Caruso, Rosario; Messina, Marianna; Maccarrone, Giuseppe; Contino, Annalinda; Cifuentes, Alejandro; Cucinotta, Vincenzo



Evaluation of a Fluorescent Lectin-Based Staining Technique for Some Acidophilic Mining Bacteria  

PubMed Central

A fluorescence-labeled wheat germ agglutinin staining technique (R. K. Sizemore et al., Appl. Environ. Microbiol. 56:2245–2247, 1990) was modified and found to be effective for staining gram-positive, acidophilic mining bacteria. Bacteria identified by others as being gram positive through 16S rRNA sequence analyses, yet clustering near the divergence of that group, stained weakly. Gram-negative bacteria did not stain. Background staining of environmental samples was negligible, and pyrite and soil particles in the samples did not interfere with the staining procedure.

Fife, Dee Jay; Bruhn, Debby F.; Miller, Karen S.; Stoner, Daphne L.



Interphase ribosomal RNA cistron staining in chronic myeloid leukaemia  

PubMed Central

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

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



Ring stains in the presence of electromagnetohydrodynamic interactions  

NASA Astrophysics Data System (ADS)

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

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



Crystal violet staining to quantify Candida adhesion to epithelial cells.  


In vitro studies of adhesion capability are essential to characterise the virulence of Candida species. However, the assessment of adhesion by traditional methods is time-consuming. The aim of the present study is the development of a simple methodology using crystal violet staining to quantify in vitro adhesion of different Candida species to epithelial cells. The experiments are performed using Candida albicans (ATCC 90028), C. glabrata (ATCC 2001), C. parapsilosis (ATCC 22019) and C. tropicalis (ATCC 750). A human urinary bladder epithelial cell line (TCC-SUP) is used. Yeast and epithelial cells were stained with crystal violet, epithelial cells were then destained using intermediate washing, and the dye in the yeast cells was extracted with acetic acid. The method was validated for the different Candida reference species by comparison with traditional microscope observation and enumeration. The method was then used to assess Candida adhesion to epithelial cells and also to silicone. For all Candida spp. high correlation values (r2= 0.9724-0.9997) between the number of adherent yeasts (microscope enumeration) and absorbance values were obtained for an inoculum concentration >10(6) cells/mL. The proposed technique was easy to perform and reproducible, enabling the determination of adhesion ability of Candida species to an epithelial cell line. PMID:20973406

Negri, M; Gonçalves, V; Silva, S; Henriques, M; Azeredo, J; Oliveira, R



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


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

Duerr, Janet S



Stained glasses under the nuclear microprobe: A window into history  

NASA Astrophysics Data System (ADS)

Stained glass fragments from the 15th, 16th and 20th centuries, belonging to Mosteiro de Santa Maria da Vitória, Batalha (Portugal), were characterised non-destructively in a nuclear microprobe. The work aimed at finding the composition of the glasses and glass paintings and relating these with the corresponding production periods. The elemental compositions of the glass fragments were obtained by means of scanning micro-beam Particle Induced X-ray Emission (?-PIXE) spectrometry in selected cross-sections. These were complemented by micro X-Ray fluorescence spectrometry. Characterisation of colour was performed by optical absorption spectroscopy in the UV-vis range, while the corrosion products were identified by optical microscopy and ?-FTIR (Fourier Transform Infra Red) spectroscopy in combination with the data generated by ?-PIXE. Nuclear microprobe analysis allowed unveiling the compositions and structures, in particular of glass paintings and corrosion products. While it is not surprising that Fe, Cu and Pb were the main elements identified in the grisaille paintings of all studied periods, as well as Ag and Cu found in the glasses decorated with yellow silver painting, their distribution gave important clues on the materials and techniques used to manufacture these stained glasses. Furthermore, it allowed establishing a definite relation between the compositions found and the periods of production, with the added bonus of correctly reassigning the manufacturing period of some samples.

Vilarigues, M.; Fernandes, P.; Alves, L. C.; da Silva, R. C.



Sequential use of the PAP and immunogold staining method for the light microscopical double staining of tissue antigens.  


Double immunoperoxidase staining using different couplers can give various combinations of colours on a single tissue section to achieve a comparable picture of different antigens. However, the colour combinations achieved to date are not entirely satisfactory. A double immunostaining procedure is introduced here, combining the peroxidase anti-peroxidase (PAP) and immunogold staining (IGS) methods. The IGS method is a new, simple, sensitive and reliable approach to immunostaining at the light microscopic level. It was carried out in three ways. Firstly, a two-step method was used in which the second layer was goat anti-rabbit IgG absorbed onto gold particles (GAR/Au20). Secondly, a three-step method was employed where the second layer was unlabelled goat anti-rabbit IgG and the third layer was a rabbit antibody to peroxidase absorbed onto the gold particles (RAP/Au20) and acting as a gold-labelled IgG antigen. The third method combined the first two methods using GAR/Au20 as th second layer and RAP/Au20 as the third layer which increased the amount of bound gold and enhanced the red colour, providing a better picture. The use of gold-labelled antibodies in double immunostaining has great potential value for many studies including that of the diffuse neuroendocrine system of the gut. PMID:7015422

Gu, J; de Mey, J; Moeremans, M; Polak, J M



A plant dye from Lawsonia inermis for protein staining after polyacrylamide gel electrophoresis.  


A reddish-brown dye was isolated from the leaves of Lawsonia inermis by extraction with calcium hydroxide (pH 11-12). A 3.6% crude extract in ethanol/water, 1:1 v/v, was used for direct staining, without fixation, of bovine serum albumin, casein and human serum proteins, following polyacrylamide gel electrophoresis in cylindrical gels. After staining for 30 min the gels were destained for 1/2-2 h with 7% acetic acid at 60 degrees C. Protein staining with Amido Black 10B and Coomassie Brilliant Blue R-250, according to standard protocols, required destaining for 24 h and more to obtain a comparably cleared background. Staining with the plant dye and Coomassie Brilliant Blue had a similar overall staining sensitivity but some minor components of human serum showed different staining characteristics with each of the two dyes. Staining with the plant dye excels over standard staining by speed and simplicity. PMID:1692790

Ali, R; Sayeed, S A



Out damn spot!--a study of the removal of dental material stains from cloth.  


Stains to white cotton caused by clinical dental materials were subjected to washing and dry cleaning processes. Most stains were successfully removed with the exception of a siloxane impression polymer. PMID:8975064

Cunningham, J L; Bolas, A



Combined silver staining of the nucleolus organizing regions and Giemsa banding in human chromosomes  

Microsoft Academic Search

A combination of the silver-staining method of the nucleolus organizer regions (NORs) with a Giemsa-banding method is deccribed. This double staining allows a rapid identification of the NOR-bearing chromosomes.

H. Zankl; S. Bernhardt



A rapid staining procedure to demonstrate glycocalyx production and bacterial biofilms.  


A novel staining procedure to demonstrate glycocalyx production by clinical isolates is presented. The short times required, specificity and sensitivity suggest that the staining could be applied to routine in vitro diagnostic procedures. PMID:7526130

Passariello, C; Berlutti, F; Selan, L; Thaller, M C; Pezzi, R



A Flow-Cytometric Gram-Staining Technique for Milk-Associated Bacteria  

Microsoft Academic Search

A Gram-staining technique combining staining with two fluorescent stains, Oregon Green-conjugated wheat germ agglutinin (WGA) and hexidium iodide (HI) followed by flow-cytometric detection is described. WGA stains gram-positive bacteria while HI binds to the DNA of all bacteria after permeabilization by EDTA and incubation at 50°C for 15 min. For WGA to bind to gram-positive bacteria ,a3M potassium chloride solution

Claus Holm; Lene Jespersen



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

Microsoft Academic Search

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

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



[Automated analysis of bacterial preparations manufactured on automatic heat fixation and staining equipment].  


Heat fixation of preparations was made in the fixation bath designed by EMKO (Russia). Programmable "Emkosteiner" (EMKO, Russia) was used for trial staining. Reagents set Micko-GRAM-NITsF was applied for Gram's method of staining. It was demostrated that automatic smear fixation equipment and programmable staining ensure high-quality imaging (1% chromaticity variation) good enough for standardization of Gram's staining of microbial preparations. PMID:23116043


Development and application of immunocytochemical staining techniques: a review.  


Since the field of immunocytochemistry was pioneered almost half a century ago, over 30 different immunostaining methods have been developed. Most laboratory investigators, however, are aware of only a few of the more popular techniques. This article, therefore, reviews and places in historical perspective many of the less common methodologies. A discussion of the progression of immunostaining methodology from purely immunologic techniques (employing only antigens, antibodies, and free enzymatic preparations) to procedures that use non-immunologic substances (such as avidin, biotin, and protein A) is presented. Basic principles of immunostaining, including specimen preparation, fixation, control procedures, interpretation, and problems unique to cytopathology, are also discussed. Schematic diagrams that illustrate the nature and arrangement of the immunochemical reagents employed in most of these techniques are provided in order to demonstrate how various materials can be combined to create a desired staining effect. PMID:2477206

Myers, J D



Sizing of single fluorescently stained DNA fragments by scanning microscopy  

PubMed Central

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

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



Ring stains in the presence of electrokinetic interactions  

NASA Astrophysics Data System (ADS)

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

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



Green staining of clothing: a signal for pseudomonal infection.  


Anticonvulsant hypersensitivity syndrome (AHS) is a nondose-related idiosyncratic reaction to aromatic antiepileptic drugs and is a cause of drug discontinuation. Pseudomonas aeruginosa is a gram-negative bacillus that can produce infections in many different organs, including the skin and soft tissue. We report a patient with erythroderma and AHS who developed a pseudomonal infection. Green staining of the underwear served as a diagnostic clue for severe P aeruginosa infection that had developed because of a local flexural skin infection that spread due to a damaged skin barrier. Inspection of the patient's clothes may give information about any exudate from the skin and should be done routinely as part of the physical examination. PMID:21284282

Yilmaz, Eylem; Savk, Ekin; Oncü, Serkan; Güleç, Güliz Uyar; Ertu?rul, Bülent; Sakarya, Serhan; Uslu, Meltem; Karaman, Göksun; Sendur, Neslihan



Preliminary oxidation in histochemical staining methods for cholesterol.  


The need for preliminary oxidation with histochemical methods for cholesterol was investigated on silica-coated sheets and in tissue sections. The techniques used were the Schultz reaction, perchloric acid-naphthoquinone (PAN), Lewis & Lobban's ferric alum-sulphuric acid reagent and Okamoto's iodine-sulphuric acid. The oxidants assessed were ferric chloride, ferric alum, potassium permanganate, ammonium sulphamate and ultraviolet light. The best combinations amongst those tested in order of reactivity were FeCl3-PAN, ferric alum-Schultz, Lewis-Lobban (no additional oxidant), iodine-sulphuric acid (no additional oxidant). Authentic preparations of cholesterol oxidation products were stained with these methods, but the nature of the oxidized product in the preliminary stage could not be determined. PMID:6157826

Adams, C W; High, O B



Method for Whole Mount Antibody Staining in Chick  

PubMed Central

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ideal tool for studying antibody expression in developing brain, neural tube and somite. This video demonstrates the different steps in whole-mount antibody staining using HRP conjugated secondary antibodies; First, the embryo is dissected from the egg and fixed in paraformaldehyde. Second, endogenous peroxidase is inactivated; The embryo is then exposed to primary antibody. After several washes, the embryo is incubated with secondary antibody conjugated to HRP. Peroxidase activity is revealed using reaction with diaminobenzidine substrate. Finally, the embryo is fixed and processed for photography and sectioning. The advantage of this method over the use of fluorescent antibodies is that embryos can be processed for wax sectioning, thus enabling the study of antigen sites in cross section. This method was originally introduced by Jane Dodd and Tom Jessell 1.

Psychoyos, Delphine; Finnell, Richard



Staining of internal limiting membrane in macular hole surgery.  


Removal of internal limiting membranes (ILMs) is a potentially useful surgical approach to close an idiopathic macular hole. However, the removal of ILMs is difficult to perform because of poor visibility of the ILMs. We have developed a technique for staining the ILM with a solution of indocyanine green to facilitate the removal of ILMs in eyes with an idiopathic macular hole. Thirteen eyes of 13 patients (8 women and 5 men, aged from 54 to 68 years) with idiopathical macular hole stage 3 or stage 4 that underwent removal of ILMs using this technique had an anatomical closure rate of 92% and an improvement of visual acuity of 89% (>/=2 Snellen letter chart lines). The excised specimens were evaluated using transmission electron microscopy. Our results show that this technique is safe and useful in visualizing the ILM, leading to the performance of successful removal of an ILM with least damage to the retina. Arch Ophthalmol. 2000;118:1116-1118 PMID:10922208

Kadonosono, K; Itoh, N; Uchio, E; Nakamura, S; Ohno, S



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.


Visualizing quantum dots in biological samples using silver staining.  


Quantum dot (QD) based contrast agents are currently being developed as probes for bioimaging and as vehicles for drug delivery. The ability to detect QDs, regardless of fluorescence brightness, in cells, tissues, and organs is imperative to their development. Traditional methods used to visualize the distribution of QDs in biological samples mainly rely on fluorescence imaging, which does not account for optically degenerate QDs as a result of oxidative quenching within the biological environment. Here, we demonstrate the use of silver staining for directly visualizing the distribution of QDs within biological samples under bright field microscopy. This strategy involves silver deposition onto the surface of QDs upon reduction by hydroquinone, effectively amplifying the size of QDs until visible for detection. The method can be used to detect non-fluorescent QDs and is fast, simple, and inexpensive. PMID:19408951

Chou, Leo Y T; Fischer, Hans C; Perrault, Steve D; Chan, Warren C W



Mechanisms To Assess Gram Stain Interpretation Proficiency of Technologists at Satellite Laboratories  

Microsoft Academic Search

To address Gram stain interpretation proficiency in a satellite\\/centralized microbiology laboratory para- digm, two programs were devised. In quality assurance program 1, nonmicrobiology technologists at satellite laboratories were required to interpret standardized Gram-stained specimens of clinical material prepared by an experienced microbiologist at a central laboratory. In quality assurance program 2, clinical Gram stains prepared and read by the satellite

Erik Munson; Timothy Block; Janice Basile; Jeanne E. Hryciuk; Ronald F. Schell


Immunocytochemical staining of breast carcinoma with the monoclonal antibody NCRC 11: a new prognostic indicator  

Microsoft Academic Search

The staining of breast cancer with a new monoclonal antibody, NCRC 11, was studied in a series of 126 women with primary breast carcinoma. Tumour samples embedded in paraffin were tested, and the minimum duration of follow up was five years or to death. Altogether 119 tumours stained positively. There was a strong relation between the intensity of staining, divided

I O Ellis; C P Hinton; J MacNay; C W Elston; A Robins; A A Owainati; R W Blamey; R W Baldwin; B Ferry



The role of intraoperative gram stain in revision total joint arthroplasty  

Microsoft Academic Search

The ability to identify intraoperatively patients with an infected prosthesis at the time of a revision procedure assists the surgeon in selecting appropriate management. The results of 413 intraoperative Gram stains were compared with the results of operative cultures, permanent histology, and the surgeon's intraoperative assessment to determine the ability of Gram stains to identify periprosthetic infection. Gram staining correctly

Craig J. Della Valle; David M. Scher; Yong H. Kim; Cubyson M. Oxley; Panna Desai; Joseph D. Zuckerman; Paul E. Di Cesare



The application of image cytometry to viability assessment in dual fluorescence-stained fish spermatozoa  

Microsoft Academic Search

The viability of spermatozoa has been assessed using SYBR 14 staining for DNA of living cells and propidium iodide staining for DNA of degenerate cells. This dual staining was performed on four fish species (Siberian sturgeon, Acipenser baerii; common carp, Cyprinus carpio; tench, Tinca tinca and wels, Silurus glanis) and the proportions of live and dead spermatozoa were assessed by

Martin Flajšhans; Jacky Cosson; Marek Rodina; Otomar Linhart



Statistical modeling, detection, and segmentation of stains in digitized fabric images  

Microsoft Academic Search

This paper will describe a novel and automated system based on a computer vision approach, for objective evaluation of stain release on cotton fabrics. Digitized color images of the stained fabrics are obtained, and the pixel values in the color and intensity planes of these images are probabilistically modeled as a Gaussian Mixture Model (GMM). Stain detection is posed as

Arunkumar Gururajan; Hamed Sari-Sarraf; Eric F. Hequet



Sudan black B as a histological stain for polymeric biomaterials embedded in glycol methacrylate.  


Sudan black B, usually a stain for all kinds of lipid, turned out to be an excellent histological stain for polymeric biomaterials embedded in glycol methacrylate. Staining the surrounding connective tissue with toluidine blue-basic fuchsin makes details of the polymer-tissue interface clearly visible. Sudan black B might be used to visualize the biodegradation process of polymeric biomaterials. PMID:3224132

Hoeksma, E A; van der Lei, B; Jonkman, M F



Evaluation of an indirect fluorescent-antibody stain for detection of Pneumocystis carinii in respiratory specimens.  

PubMed Central

Two prospective studies were undertaken to evaluate a commercial indirect fluorescent-antibody (IFA) stain for the detection of Pneumocystis carinii in respiratory specimens from individuals at risk for or with the acquired immunodeficiency syndrome. The first study compared IFA with Diff-Quik (DQ; a rapid Giemsa-like stain) for detecting P. carinii in 95 induced sputa obtained from 77 asymptomatic patients who had survived one previous episode of P. carinii pneumonia and who were being treated prophylactically with aerosolized pentamidine. Only one induced sputum specimen was found to contain P. carinii; organisms were detected by both stains. The second study compared the performance of the IFA stain versus DQ, modified toluidine blue O, and Gomori methenamine silver stains for detecting P. carinii in symptomatic individuals at risk for or with acquired immunodeficiency syndrome. Of 182 specimens examined, P. carinii was detected in 105 by one or more stains; the DQ stain detected 73 (70%), the modified toluidine blue O stain detected 75 (71%), the Gomori methenamine silver stain detected 76 (72%), and the IFA stain detected 95 (90%). The IFA stain was more sensitive (P less than 0.01) than the other traditional stains for detecting P. carinii; however, a subsequent clinical evaluation revealed that a subset of IFA-positive-only specimens were from patients whose clinical symptoms resolved without specific anti-P. carinii therapy. Images

Ng, V L; Yajko, D M; McPhaul, L W; Gartner, I; Byford, B; Goodman, C D; Nassos, P S; Sanders, C A; Howes, E L; Leoung, G



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

Microsoft Academic Search

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

S. E. Bloom; C. Goodpasture



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



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

Microsoft Academic Search

The dust on the camera's lens will leave dark stains on the image. Calibrating and compensating the intensity of the stained pixels play an important role in the airborne image processing. This article introduces an automatic compensation algorithm for the dark stains. It's based on the theory of flat-field correction. We produced a whiteboard reference image by aggregating hundreds of

Gang Wang; Dongmei Yan; Yang Yang



A comparison of constitutive heterochromatin staining methods in two cases of familial heterochromatin deficiencies  

Microsoft Academic Search

Using DAPI staining after pretreatment with distamycin A we detected a familial deficiency of chromosome 16 heterochromatin. A distinct positively staining band, however, was seen after C-banding. Thus, by using these different heterochromatin staining methods, heterogeneity of the constitutive heterochromatin in the centromeric region of human chromosome 16 was indicated. The same C-banding procedure was also applied to a previously

C. H. C. M. Buys; G. J. P. A. Anders; W. L. Gouw; J. M. M. Borkent-Ypma; J. A. M. Blenkers-Platter



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



An improved thioflavine S method for staining neurofibrillary tangles and senile plaques in Alzheimer's disease  

Microsoft Academic Search

Large differences are usually observed when standard staining methods for a number of pathological lesions in neurodegenerative disorders are compared. With the modified thioflavine S method presented here (easy and cheap to perform), the morphological appearance of the stained neurofibrillary tangles (NFT) and senile plaques (SP) is greatly improved. Furthermore, the intense contrast between stained lesions and background obtained with

R. Guntern; C. Bouras; P. R. Hof; P. G. Vallet



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


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

Bautista, Pinky A; Yagi, Yukako



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

NASA Astrophysics Data System (ADS)

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

Bautista, Pinky A.; Yagi, Yukako



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




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

PubMed Central

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

Ankle, Madhuri R; Joshi, Priya S