Sample records for fluorescein diacetate staining

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

    EPA Science Inventory

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

  2. Relationship between fluorescein diacetate-stained hyphae and oxygen utilization, glucose utilization, and biomass of submerged fungal batch cultures.

    PubMed

    Ingham, E R; Klein, D A

    1982-08-01

    The relationship between fungal activity and staining with fluorescein diacetate (FDA) was investigated by growing Penicillium citrinum and Rhizoctonia solani in submerged batch cultures at different initial glucose concentrations and aeration rates. A modified FDA staining method, similar to the Jones and Mollison technique (P. Jones and J. Mollison, J. Gen. Microbiol. 2:54-69, 1948), was developed to assess both total and FDA-stained hyphae. In previous studies, soil hyphae stained with FDA were considered viable. However, determination of a quantitative relationship between FDA staining and fungal activity is necessary before such an assumption can be made. Growth rates and the rate of change in the percentage of FDA-stained hyphae were significantly correlated. The regression equation calculated for the relationship was: growth rate (mg . ml-1 . h-1) = 0.34 + 1.1 (rate of change in the percentage of FDA-stained hyphae [. ml-1 . h-1]). Changes in activity as measured by O2 utilization, glucose utilization, and biomass correlated significantly with changes in the percentage of FDA-stained hyphae, although the relationships among these parameters were different for each fungal species. Fungal growth stage was also correlated with the percentage of FDA-stained hyphae. Staining was 10% or greater during fungal growth and less than 10% during the late growth, stationary, and death phases. Thus, the rate of change in the percentage of FDA-stained hyphae can be used to predict fungal activity rate changes for single fungal cultures and growth rates for mixed fungal cultures, and the growth stage can be assessed by the percentage of FDA-stained hyphae. PMID:7125653

  3. Relationship between fluorescein diacetate-stained hyphae and oxygen utilization, glucose utilization, and biomass of submerged fungal batch cultures.

    PubMed Central

    Ingham, E R; Klein, D A

    1982-01-01

    The relationship between fungal activity and staining with fluorescein diacetate (FDA) was investigated by growing Penicillium citrinum and Rhizoctonia solani in submerged batch cultures at different initial glucose concentrations and aeration rates. A modified FDA staining method, similar to the Jones and Mollison technique (P. Jones and J. Mollison, J. Gen. Microbiol. 2:54-69, 1948), was developed to assess both total and FDA-stained hyphae. In previous studies, soil hyphae stained with FDA were considered viable. However, determination of a quantitative relationship between FDA staining and fungal activity is necessary before such an assumption can be made. Growth rates and the rate of change in the percentage of FDA-stained hyphae were significantly correlated. The regression equation calculated for the relationship was: growth rate (mg . ml-1 . h-1) = 0.34 + 1.1 (rate of change in the percentage of FDA-stained hyphae [. ml-1 . h-1]). Changes in activity as measured by O2 utilization, glucose utilization, and biomass correlated significantly with changes in the percentage of FDA-stained hyphae, although the relationships among these parameters were different for each fungal species. Fungal growth stage was also correlated with the percentage of FDA-stained hyphae. Staining was 10% or greater during fungal growth and less than 10% during the late growth, stationary, and death phases. Thus, the rate of change in the percentage of FDA-stained hyphae can be used to predict fungal activity rate changes for single fungal cultures and growth rates for mixed fungal cultures, and the growth stage can be assessed by the percentage of FDA-stained hyphae. PMID:7125653

  4. 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

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

    2010-01-01

    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. PMID:20814586

  5. Fluorescein eye stain

    MedlinePLUS

    ... test that uses orange dye (fluorescein) and a blue light to detect foreign bodies in the eye. This ... eye. The health care provider then shines a blue light at your eye. Any problems on the surface ...

  6. ASSAY FOR FLUORESCEIN DIACETATE HYDROLYTIC ACTIVITY: OPTIMIZATION FOR SOIL SAMPLES

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    Microsoft Academic Search

    Ronald Swisher; George C. Carroll

    1980-01-01

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

  8. Fluorescein diacetate hydrolysis as a measure of fungal biomass in soil.

    PubMed

    Gaspar, M L; Cabello, M N; Pollero, R; Aon, M A

    2001-05-01

    The fatty acid methyl esters of lipids extracted from an agricultural soil in the preharvest period of soybean or middle growth cycle from wheat were characterized and quantified by gas-liquid chromatography. The fatty acids 18:2omega6 and 16:1omega5 were used as markers of saprotrophic and arbuscular mycorrhizal fungi. In parallel, biomass estimation through plate counts in selective media for cellulolytic and saprotrophic fungi was also performed all throughout a soybean crop or middle growth cycle of wheat. As an enzymatic method, the fluorescein diacetate (FDA) hydrolytic activity of the samples was determined. Owing to the high relationship exhibited by FDA hydrolysis with organic carbon and total nitrogen content of soil, the enzymatic activity was correlated with the microbial biomass estimated through marker lipids or plate counts. The results obtained point out that FDA hydrolysis may be used as a rapid, cheap, and reliable estimator of fungal biomass. PMID:11400054

  9. STAINING TOXOPLASMA GONDII WITH FLUORESCEIN-LABELLED ANTIBODY

    PubMed Central

    Goldman, Morris

    1957-01-01

    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. PMID:13428924

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

    PubMed Central

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

    2012-01-01

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

  11. Development of an automated ballast water treatment verification system utilizing fluorescein diacetate hydrolysis as a measure of treatment efficacy.

    PubMed

    Akram, A C; Noman, S; Moniri-Javid, R; Gizicki, J P; Reed, E A; Singh, S B; Basu, A S; Banno, F; Fujimoto, M; Ram, J L

    2015-03-01

    Methods for verifying ballast water treatments in foreign vessels are needed to protect the Great Lakes from the discharge of live non-native organisms or pathogens. A prototype automated viability test system using fluorescein diacetate (FDA), a membrane permeable fluorogen, to differentiate live from dead bacteria and algae is described. The automated fluorescence intensity detection device (AFIDD) captures cultured algae or organisms in Detroit River water (simulated ballast water) on 0.2 ?m filters, backwashes them from the filter into a cuvette with buffer and FDA for subsequent fluorescence intensity measurements, and washes the filters with sterile water for serial automated reuse. Preliminary manual versions of these procedures were also tested. Tests of various buffers determined N,N-Bis(2-hydroxyethyl)-2-aminoethanesulfonic acid, N,N-Bis(2-hydroxyethyl)taurine (BES) and 3-(N-morpholino)propanesulfonic acid (MOPS) at pH 7.0 to be the best buffers, causing the least spontaneous FDA breakdown without inhibiting enzymatic activity. Fluorescence in the presence of live organisms increased linearly over time, and the rate of increase was dependent on the sample concentration. Following simulated ballast water treatments with heat or chlorine, the fluorescence produced by Detroit River samples decreased to near control (sterile water) levels. Automated measurements of FDA hydrolysis with a reusable filter backwash system should be applicable to near real-time remote-controlled monitoring of live organisms in ballast water. PMID:25555225

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

    PubMed Central

    Tabery, H M

    1992-01-01

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

  13. Fluorescein. Physiochemical factors affecting its fluorescence.

    PubMed

    Romanchuk, K G

    1982-01-01

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

  14. 21 CFR 582.6197 - Calcium diacetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

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

  15. 21 CFR 582.6197 - Calcium diacetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

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

  16. 21 CFR 182.6197 - Calcium diacetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

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

  17. 21 CFR 182.6197 - Calcium diacetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

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

  18. 21 CFR 182.6197 - Calcium diacetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

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

  19. 21 CFR 182.6197 - Calcium diacetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

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

  20. 21 CFR 182.6197 - Calcium diacetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

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

  1. 21 CFR 582.6197 - Calcium diacetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

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

  2. 21 CFR 582.6197 - Calcium diacetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

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

  3. 21 CFR 582.6197 - Calcium diacetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

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

  4. 21 CFR 582.6754 - Sodium diacetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 2010-04-01 false Sodium diacetate. 582.6754 Section 582...SAFE Sequestrants 2 § 582.6754 Sodium diacetate. (a) Product. Sodium diacetate. (b) Conditions of use. This substance is generally...

  5. 21 CFR 582.6754 - Sodium diacetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...2011-04-01 2011-04-01 false Sodium diacetate. 582.6754 Section 582...SAFE Sequestrants 2 § 582.6754 Sodium diacetate. (a) Product. Sodium diacetate. (b) Conditions of use. This substance is generally...

  6. 21 CFR 582.6754 - Sodium diacetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...2013-04-01 2013-04-01 false Sodium diacetate. 582.6754 Section 582...SAFE Sequestrants 2 § 582.6754 Sodium diacetate. (a) Product. Sodium diacetate. (b) Conditions of use. This substance is generally...

  7. 21 CFR 582.6754 - Sodium diacetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...2014-04-01 2014-04-01 false Sodium diacetate. 582.6754 Section 582...SAFE Sequestrants 2 § 582.6754 Sodium diacetate. (a) Product. Sodium diacetate. (b) Conditions of use. This substance is generally...

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

    PubMed Central

    2014-01-01

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

  9. Fluorescein Derivatives in Intravital Fluorescence Imaging

    PubMed Central

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

    2013-01-01

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

  10. Fluorescein angiography findings in phacomatosis pigmentovascularis.

    PubMed

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

    2013-01-01

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

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

    PubMed Central

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

    2014-01-01

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

  12. Gram stain

    MedlinePLUS

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

  13. 21 CFR 184.1754 - Sodium diacetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1754 Sodium diacetate. (a) Sodium diacetate (C4 H7 O4 Na·xH2 O, CAS Reg. No. 126-96-5) is a molecular compound of acetic acid, sodium acetate, and...

  14. Fluorescein-related extensive jaundice.

    PubMed

    Kalkan, Asim; Turedi, Suleyman; Aydin, Ibrahim

    2015-03-01

    Fluorescein is a chemical dye frequently used in eye diseases to assess blood flow in the retina, choroid tissue, and iris. Although it has many known adverse effects, it has not previously been reported to lead to jaundice. The purpose of this case report was to emphasize that for patients presenting at the emergency department with jaundice symptoms, it should not be forgotten by emergency physicians that jaundice can develop after fluorescein angiography. Case: A 65-year-old woman presented at the emergency department with extensive jaundice that had developed on her entire body a few hours after fluorescein angiography applied because of vision impairment. The test results for all the diseases considered to cause jaundice were normal,and fluorescein-related jaundice was diagnosed. Conclusion: A detailed anamnesis should be taken when jaundice is seen in patients who have undergone fluorescein angiography, and it should not be forgotten that fluorescein dye is a rare cause of jaundice. PMID:25245286

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

    Microsoft Academic Search

    Y Yang; S Moon; S Lee; J Kim

    1996-01-01

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

  16. Seidel's test using 10% fluorescein.

    PubMed

    Romanchuk, K G

    1979-10-01

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

  17. Study of the biodistribution of fluorescein in glioma-infiltrated mouse brain and histopathological correlation of intraoperative findings in high-grade gliomas resected under fluorescein fluorescence guidance.

    PubMed

    Diaz, Roberto Jose; Dios, Roberto Rey; Hattab, Eyas M; Burrell, Kelly; Rakopoulos, Patricia; Sabha, Nesrin; Hawkins, Cynthia; Zadeh, Gelareh; Rutka, James T; Cohen-Gadol, Aaron A

    2015-06-01

    OBJECT Intravenous fluorescein sodium has been used during resection of high-grade gliomas to help the surgeon visualize tumor margins. Several studies have reported improved rates of gross-total resection (GTR) using high doses of fluorescein sodium under white light. The recent introduction of a fluorescein-specific camera that allows for high-quality intraoperative imaging and use of very low dose fluorescein has drawn new attention to this fluorophore. However, the ability of fluorescein to specifically stain glioma cells is not yet well understood. METHODS The authors designed an in vitro model to assess fluorescein uptake in normal human astrocytes and U251 malignant glioma cells. An in vivo experiment was also subsequently designed to study fluorescein uptake by intracranial U87 malignant glioma xenografts in male nonobese diabetic/severe combined immunodeficient mice. A genetically induced mouse glioma model was used to adjust for the possible confounding effect of an inflammatory response in the xenograft model. To assess the intraoperative application of this technology, the authors prospectively enrolled 12 patients who underwent fluorescein-guided resection of their high-grade gliomas using low-dose intravenous fluorescein and a microscope-integrated fluorescence module. Intraoperative fluorescent and nonfluorescent specimens at the tumor margins were randomly analyzed for histopathological correlation. RESULTS The in vitro and in vivo models suggest that fluorescein demarcation of glioma-invaded brain is the result of distribution of fluorescein into the extracellular space, most likely as a result of an abnormal blood-brain barrier. Glioblastoma tumor cell-specific uptake of fluorescein was not observed, and tumor cells appeared to mostly exclude fluorescein. For the 12 patients who underwent resection of their high-grade gliomas, the histopathological analysis of the resected specimens at the tumor margin confirmed the intraoperative fluorescent findings. Fluorescein fluorescence was highly specific (up to 90.9%) while its sensitivity was 82.2%. False negatives occurred due to lack of fluorescence in areas of diffuse, low-density cellular infiltration. Margins of contrast enhancement based on intraoperative MRI-guided StealthStation neuronavigation correlated well with fluorescent tumor margins. GTR of the contrast-enhancing area as guided by the fluorescent signal was achieved in 100% of cases based on postoperative MRI. CONCLUSIONS Fluorescein sodium does not appear to selectively accumulate in astrocytoma cells but in extracellular tumor cell-rich locations, suggesting that fluorescein is a marker for areas of compromised blood-brain barrier within high-grade astrocytoma. Fluorescein fluorescence appears to correlate intraoperatively with the areas of MR enhancement, thus representing a practical tool to help the surgeon achieve GTR of the enhancing tumor regions. PMID:25839919

  18. Stain Reagent Reversible Stain Kits

    E-print Network

    Lebendiker, Mario

    the protein is stained, not the gel, allowing protein bands to be viewed directly within the staining reagent during the staining process. 1 Water Wash-Enhanced Protein Staining With GelCode ® Blue Stain Reagent Introduction Coomassie® Brilliant Blue is the most common dye used to stain proteins on polyacrylamide gels

  19. "Geyser" leakage on fluorescein angiography.

    PubMed

    Levy, Jaime; Fagan, Xavier J; Lifshitz, Tova; Schneck, Marina

    2013-01-01

    An 82-year-old patient with diabetes was followed up due to moderate nonproliferative diabetic retinopathy with macular edema in the right eye. Visual acuity was 6/36. Focal macular laser was conducted (A). Three years later, the patient presented with blurry vision in the right eye. Visual acuity was 3/60. Vitreous hemorrhage was observed (B), and neovascularization of the disc was suspected (C). Fluorescein angiography (D, mid venous phase; E-F, recirculation phase) confirmed neovascularization of the disc and depicted a striking vertical leakage. Panretinal photocoagulation was started. Possible explanations for the "geyser" leakage may be either a partial posterior vitreous detachment allowing the fluorescein to track upwards but not elsewhere or a pocket of syneretic vitreous allowing the fluorescein passage in which to diffuse, much like the passage the blood would have taken. PMID:24548789

  20. FLUORESCEIN-CONJUGATED BOVINE ALBUMIN

    PubMed Central

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

    1953-01-01

    Fluorescein-bovine albumin conjugates have been prepared and found not to differ appreciably in size, shape, and homogeneity from the precursor, bovine serum albumin. Fluorescein has also been conjugated to rat plasma proteins. Their disappearance rates from the circulation of rats correspond with those obtained from the use of isotope labeling. Their sites of localization in rat tissues were shown to be in the cytoplasm but not in the nuclei of Kupffer cells, fixed macrophages, granulocytes, and proximal renal tubules. Adsorption to endothelium was a characteristic finding. Extracellular localizations were predominantly in the lumina of blood vessels and proximal renal tubules (but never in the lumina of collecting tubules), and the interstitial fluid of skeletal and cardiac muscle (but not that of glandular organs such as the adrenals, liver, and spleen). BAC absorption from the skin of rabbits requires days whereas sodium fluorescein absorption is measured in hours, attesting to the persistence of the colloidal state of BAC in vivo. Fluorescein conjugates have been used to visualize the transcapillary passage of circulating proteins in the mesenteric circulation of frogs and rats by direct microscopic observation and found to diffuse slowly in the manner predicted for plasma proteins. The normal cutaneous vessels of the rat are impermeable in the gross to the labeled proteins; second degree burn promptly increases the permeability of these vessels rendering the presence of the label detectable in the gross in the skin. The process of labeling does not render guinea pig albumin antigenic, although slight antigenicity results from labeling whole plasma protein. It is believed that sufficient biological evidence is presented to support the conclusion that fluorescein-conjugated plasma proteins, particularly albumin, behave in vivo like their native precursors. PMID:13035065

  1. Method and compositions for detecting of bloodstains using fluorescin-fluorescein reaction

    DOEpatents

    Di Benedetto, John; Kyle, Kevin; Boan, Terry; Marie, Charlene

    2004-02-17

    A method, compositions and kit are set forth for detecting blood stains. A reactant solution includes fluorescin solubilized (reduced) in acetic acid in ethanol. The solution may be buffered to a pH of approximately 9. After spraying the reactant solution on the suspected area an oxidizer is applied to promote the fluorescin to fluorescein reaction with the blood. The reacted fluorescein is then detected through luminescence for capture by photography.

  2. Clinical grading of corneal staining of non-contact lens wearers

    Microsoft Academic Search

    Morven Dundas; Alyson Walker; Russell L. Woods

    2001-01-01

    Summary To distinguish normal from pathological corneal fluorescein staining requires knowledge of background levels of staining among otherwise healthy individuals. Corneal staining of 102 non- contact lens wearing subjects was assessed using a photographic grading scale that uses a generic (0 to 4) scale to score corneal staining. Some degree of corneal staining was found on 79% of the corneas.

  3. 76 FR 75886 - Determination That DEMULEN 1/50-28 (Ethinyl Estradiol; Ethynodiol Diacetate) Tablet and Four...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-05

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

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

    Microsoft Academic Search

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

    1999-01-01

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

  5. Report on the presence of a toxic substance, dimethyl formamide, in sodium fluorescein used for fluorescein angiography

    Microsoft Academic Search

    J S Jacob; E S Rosen; E Young

    1982-01-01

    The revelation that intravenous sodium fluorescein is not all that it might seem to be may be a significant finding in the light of the adverse reactions to fluorescein that have been previously reported. Analysis of commercially prepared intravenous sodium fluorescein by mass spectroscopy and nuclear magnetic resonance has indicated that an industrial solvent used in the manufacturing process has

  6. Joint fluid Gram stain

    MedlinePLUS

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

  7. Automated Slide Staining Machine

    PubMed Central

    Drew, W. Lawrence; Pedersen, Anders N.; Roy, Jacques J.

    1972-01-01

    A machine is described which can perform the Gram stain. Comparison of slides stained by machine versus hand revealed no difference in reproducibility or accuracy. In addition to providing clean, dry, uniformly stained slides, the machine saves 24 sec per slide when compared with a hand staining technique. Images PMID:4110426

  8. Port-Wine Stains

    MedlinePLUS

    ... port-wine stains called a "pulsed-dye" laser . Laser treatment is often started in infancy when the stain ... easier to treat. But that doesn't mean laser treatments can't help older kids or teens, too — ...

  9. Hen Fluorescein-Labeled Gonococcal Lipopolysaccharide Antibody in the Delayed Fluorescent Antibody Technique for the Confirmation of Neisseria gonorrhoeae

    PubMed Central

    Ashton, F. E.; Leitch, R. A.; Perry, M. B.; Wallace, R.; Diena, B. B.

    1979-01-01

    A fluorescent antibody reagent (termed anti-LPS conjugate) was prepared from sera obtained from hens immunized with gonococcal R-type lipopolysaccharide. The reagent was absorbed with Formalin-treated cells of Neisseria meningitidis. The anti-LPS conjugate gave uniform brilliant staining of Neisseria gonorrhoeae with little background fluorescence, thus making interpretation and reading of fluorescence simple. The conjugate did not significantly stain cultures of N. meningitidis, Neisseria lactamica, nonpathogenic Neisseria species, or other gram-negative bacteria. Several preparations of the conjugate provided the same specificity and reproducibility of staining. The anti-LPS conjugate was compared with Difco Laboratories fluorescent antibody conjugate for staining of N. gonorrhoeae. Both conjugates stained cells of the light and dark variants of gonococcal colony types 1 and 2, as well as cells of colony types 3 and 4. When used for the confirmation of N. gonorrhoeae, the anti-LPS and Difco conjugates stained 426 of 431 (98.8%) and 210 of 213 (98.6%) of the gonococcal cultures, respectively. Absorption of the anti-LPS conjugate with R-type lipopolysaccharide removed the staining of gonococci. However, absorption of Difco conjugate with R-type lipopolysaccharide did not remove the staining of gonococci, suggesting that the majority of fluorescein-labeled antibody present in the Difco conjugate is directed to gonococcal cell surface components other than lipopolysaccharide. The results of this study indicate that fluorescein-labeled gonococcal lipopolysaccharide antibody should be a reliable fluorescent antibody reagent for the confirmation of N. gonorrhoeae. PMID:110824

  10. Gram stain of urethral discharge

    MedlinePLUS

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

  11. Fluorescent staining of gels.

    PubMed

    Buxbaum, Engelbert

    2012-01-01

    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

  12. Port-wine stain

    MedlinePLUS

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

  13. Immunofluorescence analysis of cytokeratin 8/18 staining is a sensitive assay for the detection of cell apoptosis

    PubMed Central

    DONG, QIAO-MEI; LING, CHUN; ZHAO, LI

    2015-01-01

    Apoptosis is one of the major types of programmed cell death. During this process, cells experience a series of morphological and biochemical changes. Flow cytometric analysis of Annexin V staining of cell surface phosphatidylserine, in combination with a DNA-staining dye to probe the permeability of the cell membrane, is an established method for detecting apoptosis. The present study aimed to show that the immunofluorescence analysis of cytokeratin (CK) 8/18 staining may provide a new and sensitive assay for the detection of apoptotic cells. Tumor cells were treated with 20 ?M cisplatin to induce apoptosis. Following 12 and 24 h of cisplatin treatment, cells were collected and stained with 4?,6-diamidine-2?-phenylindole dihydrochloride (DAPI) and fluorescein-labeled anti-CK8/18 antibody. The apoptotic cells were subsequently examined by fluorescence microscopy. Annexin V-fluorescein isothiocyanate/propidium iodide staining followed by flow cytometric analysis confirmed that cisplatin was able to induce apoptosis in tumor cells. Immunofluorescence analysis demonstrated that apoptotic cells had a distinct CK8/18 staining pattern. In living cells, CK8/18 was uniformly distributed in the cytoplasm and cytosol; however in the apoptotic cells with a condensed and/or fragmented apoptotic nucleus (as identified by DAPI staining), fluorescein-labeled anti-CK8/18 antibody exhibited unusual punctate and/or bubbly staining in the cytosol. In the apoptotic cells that could not be identified by DAPI staining, fluorescein-labeled CK8/18 displayed polarized aggregated staining in the cytosol. These results indicate that fluorescein-conjugated CK8/18 may be a useful and sensitive indicator of cell apoptosis. PMID:25663887

  14. Fluorescent Sensors for Zn2+ Based on a Fluorescein Platform

    E-print Network

    Tsien, Roger Y.

    Fluorescent Sensors for Zn2+ Based on a Fluorescein Platform: Synthesis, Properties to the sensor, which inhibits a photoinduced electron transfer (PET) quenching pathway. The X-ray crystal properties § Massachusetts Institute of Technology. Department of Pharmacology, University of California

  15. Acute Toxicity of Sodium Fluorescein to Ashy Pebblesnails Fluminicola fuscus

    USGS Publications Warehouse

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

    2011-01-01

    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.

  16. Activity staining method of chitinase on chitin agar plate through polyacrylamide gel electrophoresis

    Microsoft Academic Search

    Vipul Gohel; Pranav Vyas; H. S. Chhatpar

    2005-01-01

    A method for detection of chitinase activity on chitin agar plate after polyacrylamide gel electrophoresis is described. Different staining dyes such as calcofluor white M2R, fluorescein isothiocyanate, rhodamine B, ruthenium red and congo red were separately incorporated in chitin agar plates. After running polyacrylamide gel electrophoresis, the gel was transferred onto chitin agar plate containing different dyes for the activity

  17. Nanoparticle Stained Glass

    NSDL National Science Digital Library

    2014-06-18

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

  18. Outward transport of fluorescein from the vitreous in aphakic eyes.

    PubMed Central

    Miyake, K; Miyake, T; Miyake, C; Asakura, M; Maekubo, K

    1985-01-01

    By administering fluorescein intravenously to 95 patients we calculated the ratio of fluorescein concentration in the vitreous at the time of its peak level compared with the estimated unbound concentration of fluorescein in the plasma at the same time. We studied 12 normal phakic and 83 aphakic eye approximately two months, one year, and more than two years after cataract extraction. All the eyes had undergone intracapsular cataract extraction or extracapsular cataract extraction, with or without posterior capsulotomy, because of senile cataract. The calculated ratio in patients with intracapsular and extracapsular lens extraction was statistically significantly reduced at two months and one year after cataract extraction and was normalised at more than two years after the operation in comparison with normal subjects. The ratio was statistically extracapsular extraction at two months and one year after surgery. Posterior capsulotomy had no effect on the ratio. The ratio, we considered, at least partially reflects the outward transport of fluorescein from the vitreous cavity. Although the findings reflect subclinical phenomena, they are of importance when considering postoperative sequelae. The posterior lens capsule, zonule, and intact anterior vitreous face may be essential for the anterior uvea to function in the outward transport of fluorescein from the vitreous cavity. PMID:4005210

  19. Pleural fluid Gram stain

    MedlinePLUS

    Gram stain of pleural fluid ... lungs fill a person's chest with air. If fluid builds up in the space outside the lungs ... chest, it can cause many problems. Removing the fluid can relieve a person's breathing problems and help ...

  20. A Model for Tear Film Thinning With Osmolarity and Fluorescein

    PubMed Central

    Braun, Richard J.; Gewecke, Nicholas R.; Begley, Carolyn G.; King-Smith, P. Ewen; Siddique, Javed I.

    2014-01-01

    Purpose. We developed a mathematical model predicting dynamic changes in fluorescent intensity during tear film thinning in either dilute or quenching regimes and we model concomitant changes in tear film osmolarity. Methods. We solved a mathematical model for the thickness, osmolarity, fluorescein concentration, and fluorescent intensity as a function of time, assuming a flat and spatially uniform tear film. Results. The tear film thins to a steady-state value that depends on the relative importance of the rates of evaporation and osmotic supply, and the resulting increase of osmolarity and fluorescein concentrations are calculated. Depending on the initial thickness, the rate of osmotic supply and the tear film thinning rate, the osmolarity increase may be modest or it may increase by as much as a factor of eight or more from isosmotic levels. Regarding fluorescent intensity, the quenching regime occurs for initial concentrations at or above the critical fluorescein concentration where efficiency dominates, while lower concentrations show little change in fluorescence with tear film thinning. Conclusions. Our model underscores the importance of using fluorescein concentrations at or near the critical concentration clinically so that quenching reflects tear film thinning and breakup. In addition, the model predicts that, depending on tear film and osmotic factors, the osmolarity within the corneal compartment of the tear film may increase markedly during tear film thinning, well above levels that cause marked discomfort. PMID:24458153

  1. Scanning laser ophthalmoscope imaging of fluorescein-labelled blood cells

    Microsoft Academic Search

    Jean-François Le Gargasson; Michel Paques; Jean-Eric Guez; Bernadette Boval; Eric Vicaut; Xin Hou; Yvon Grall; Alain Gaudric

    1997-01-01

    • Purpose: To demonstrate the feasibility of a technique for the visualization by scanning laser ophthalmoscope (SLO) of fluores cein-labelled autologous leukocytes and platelets in retinal vessels. • Method: Individual blood samples from rats and rabbits were centrifuged to isolate platelets and leukocytes, then passively labelled with fluorescein and reinjected into the same animal. An SLO was used to visualize

  2. Spectrum of complications in the use of intrathecal fluorescein.

    PubMed

    Moseley, J I; Carton, C A; Stern, W E

    1978-05-01

    The authors describe a case in which 0.5 cc of 5% fluorescein diluted in 10 cc of cerebrospinal fluid (CSF) was injected at the L4-5 level for evaluation of nasal CSF leakage. Within minutes, tone increased in lower extremities accompanied by knee and ankle clonus and subjective numbness up to the waist. Non-preserved saline irrigation of the lumbar CSF was administered until it became clear, and the patient's head was elevated to retard the developing symptomatology. Although a transient temperature elevation was observed with negative CSF cultures, all signs and symptoms cleared within 48 hours. In a survey of the members of the Americal Association of Neurological Surgeons regarding frequency of use and complications stemming from intrathecal fluorescein, the response rate was 58.3% (1111) of the 1907 members, of which 6.8% (76) had used intrathecal fluorescein, and among those, 25% (19 of the 76) had observed complications involving lower extremity weakness, numbness, generalized seizure activity, opisthotonos, and cranial nerve deficit. No complications were permanent. The authors recommend caution if intrathecal fluorescein must be used. Means should be available to clear the CSF of the agent and elevate the head if complications arise. PMID:580435

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

    SciTech Connect

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

    2012-11-09

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

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

    PubMed

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

    2014-01-01

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

  5. "Stained Glass" Landscape Windows

    ERIC Educational Resources Information Center

    Vannata, Janine

    2008-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2011-04-01

    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.

  7. Ionic Liquid-Based Fluorescein Colorimetric pH Nanosensors

    PubMed Central

    Das, Susmita; Magut, Paul K. S.; de Rooy, Sergio L.; Hasan, Farhana; Warner, Isiah M.

    2014-01-01

    A novel pH sensitive, colorimetric ionic liquid nanosensor based on phosphonium salts of fluorescein is reported. Herein, fluorescein salts of various stoichiometries were synthesized by use of a trihexyltetradecylphosphonium cation [TTP]+ in combination with dianionic [FL]2? and monoanionic [FL]? fluorescein. Nanomaterials derived from these two compounds yielded contrasting colorimetric responses in neutral and acidic environments. Variations in fluorescence spectra as a function of pH were also observed. Examination of TEM and DLS data revealed significant expansion in the diameter of [TTP]2[FL] nanodroplets in acidic environments of variable pHs. A similar trend was also observed for [TTP][FL] nanoparticles. The pH dependent colorimetric and other optical properties of these nanomaterials are attributed to alterations in molecular orientations and stacking as suggested by measuring the absorption, fluorescence, and zeta potential. Since the pH is an important indicator for many diseases, including cancer, these nanosensors are considered to be potential candidates for biomedical applications. PMID:25264488

  8. Segmentation of choroidal neovascularization in fundus fluorescein angiograms.

    PubMed

    Abdelmoula, Walid M; Shah, Syed M; Fahmy, Ahmed S

    2013-05-01

    Choroidal neovascularization (CNV) is a common manifestation of age-related macular degeneration (AMD). It is characterized by the growth of abnormal blood vessels in the choroidal layer causing blurring and deterioration of the vision. In late stages, these abnormal vessels can rupture the retinal layers causing complete loss of vision at the affected regions. Determining the CNV size and type in fluorescein angiograms is required for proper treatment and prognosis of the disease. Computer-aided methods for CNV segmentation is needed not only to reduce the burden of manual segmentation but also to reduce inter- and intraobserver variability. In this paper, we present a framework for segmenting CNV lesions based on parametric modeling of the intensity variation in fundus fluorescein angiograms. First, a novel model is proposed to describe the temporal intensity variation at each pixel in image sequences acquired by fluorescein angiography. The set of model parameters at each pixel are used to segment the image into regions of homogeneous parameters. Preliminary results on datasets from 21 patients with Wet-AMD show the potential of the method to segment CNV lesions in close agreement with the manual segmentation. PMID:23314765

  9. Quick Stain Removal Guide 

    E-print Network

    Brown, Pamela J.

    1998-07-29

    . Avoid using a rubbing motion. Launder the whole item after treating. Laundry products Soaps are mild cleansers that come in granules, which are used for lightly soiled and delicate items, or bars, which are good for pretreating heavy soils... and stains before laundering. Avoid harsh rubbing with the bar. L-5199 3-98 *Extension Consumer Science Specialist; The Texas A&M University System Pamela J. Brown* You can prevent many of these problems if you: ? Empty all pockets and close zippers...

  10. Blood stain pattern analysis

    Microsoft Academic Search

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

    2011-01-01

    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

  11. 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...

  12. Removing Stains from Washable Fabrics. 

    E-print Network

    Beard, Ann Vanderpoorten

    1988-01-01

    , then rinse. If color or texture is affected, do not use this product to treat the stain. ? When treating a spot, place it face down on white paper towels or a soft, clean, white cloth pad. Apply stain remover to the wrong side of the stain so... that the stain will be forced off the surface and not through the fabric. An eyedropper is useful for applying removers. Replace the towels or cloth pad frequently to prevent the stain from transfer ring back onto the fabric. ? Sponge by applying stain...

  13. 7.G Stained Glass

    NSDL National Science Digital Library

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

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

    SciTech Connect

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

    1980-01-01

    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.

  15. Length of stain dosimeter

    NASA Technical Reports Server (NTRS)

    Lueck, Dale E. (inventor)

    1994-01-01

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

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

    PubMed

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

    2014-01-01

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

  17. Ground beef shelf life assessment as influenced by sodium lactate, sodium propionate, sodium diacetate, and soy protein concentrate 

    E-print Network

    Grones, Kelly Leann

    2000-01-01

    cooked beef/brothy, cooked beef/fat and soda flavor and decreased cardboard and soured flavors. Sodium diacetate increased soured flavor, sour basic taste and decreased the positive effects of sodium lactate on cardboard flavor. The addition...

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

    Microsoft Academic Search

    Satoshi Koyano; Makoto Araie; Shuichiro Eguchi

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

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

    NASA Astrophysics Data System (ADS)

    Suitchmezian, Viktor; Jeß, Inke; Näther, Christian

    2006-11-01

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

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

    PubMed

    Norn, M

    1985-12-01

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

  1. A novel protocol of whole mount electro-immunofluorescence staining

    PubMed Central

    Liu, Hongshan

    2009-01-01

    Purpose To develop a new method of whole mount immunostaining that improves the penetration of staining reagents into the cornea and decreases non-specific binding and background. Methods Adult mouse corneas were fixed overnight in 4% paraformaldehyde or a mixture of 4% paraformaldehyde and 0.2% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4, at 4 °C. After washing with 0.1% Triton X-100, corneas were embedded in 1% solidified agarose in a plastic column and fluorescent staining reagents, e.g., FITC-IgG (Fluorescein isothiocyanate- immunoglobulinG) conjugates in 0.5% solidified agarose was overlaid onto the specimens. The column was directionally immersed in a submarine gel electrophoresis apparatus filled with Tris-glycine buffer (TGB, pH=7.4) and electrophoresed at 4–10 mA for 10–24 h. For comparison, conventional protocols of immune fluorescent staining were also employed. The outcomes were evaluated by confocal microscopy. Results Antibody conjugates recognizing extracellular matrix (ECM) components, integral membrane protein, and intracellular structural proteins were used in whole mount corneas. The images of confocal laser scanning microscopy (CLSM) displayed a uniform distribution pattern of keratocan in corneal stroma, which is similar to that of section-staining. Anti-?-tubulin antibodies bound to microtubes that are distributed within the whole cell body of superficial corneal epithelium cells and stromal keratocytes, but it was found perinuclear of corneal epithelial wing layers and endothelium; integral membrane protein, FAK (focal adhesion kinase), specifically labeled stromal cells of keratectomy corneas that healed for three weeks. In comparison, conventional protocols of immune fluorescent staining using the same antibody conjugates were also employed but did not yield satisfactory results. It was found that IgG conjugates examined did not readily penetrate into stroma and/or intact corneal epithelium. Phalloidin is a small molecule that can readily penetrate into deep tissue and preferentially binds to F-actin. After the whole mount electrofluorescent staining of phalloidin-rhodamine in the mouse cornea, the results were the same as conventional whole mount staining during the healing of epithelial debridement. The cytoplasmic protrusion formed by lamellipodia and filopodia can be clearly demonstrated. Conclusions These results indicate that the whole mount electro-immunofluorescent staining allows the detection of antigens in all layers of cornea, i.e., epithelium, stroma, and endothelium. PMID:19262742

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

    PubMed Central

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

    2000-01-01

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

  3. Silver stain for electron microscopy

    NASA Technical Reports Server (NTRS)

    Corbett, R. L.

    1972-01-01

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

  4. Staining Associated with Oxhorn Bucida (\\

    Microsoft Academic Search

    DOUGLAS L. CALDWELL

    ADDITIONAL INDEX WORDS. Characoma nilotica, Eriophyes, caterpillars, Combretaceae, Noctuidae, galls Oxhorn bucida (Bucida buceras L.), also known as black olive, is used as a shade tree in southern Florida landscapes and street plantings. One negative aspect associated with this tree is a rusty staining of driveways and other objects beneath the canopy. This report documents that the objectionable staining is

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

    E-print Network

    Southern California, University of

    CNT Uptake in Human Ovarian Cancer Cells: Changes in Fluorescence Emission of CNT-linked carbon nanotube (CNT)- fluorescein conjugates into SKOV-3 human ovarian cancer cells. The uptake of CNTs loaded with amide-linked CNT-fluorescein conjugates increases after brief exposure to light

  6. Removing Stains from Washable Fabrics.

    E-print Network

    Beard, Ann Vanderpoorten

    1988-01-01

    ) . are set by alkalies such as ammonia; others (blood and vomit) are set by alcohol. Avoid using chemi cals unless you are certain they will work on the stain you are treating. ? Never mix stain removers, especially ammonia and bleach. If more than one... of a ring. Use small amounts of re mover with quick light strokes. ? Flush the stain away by adding remover slowly to the spot from the wrong side of the fabric. ? When using any bleach, do not try to bleach just a spot on a colored garment. Bleach...

  7. Heterogeneous catalyst for the production of ethylidene diacetate from acetic anhydride

    DOEpatents

    Ramprasad, D.; Waller, F.J.

    1998-06-16

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

  8. Heterogeneous catalyst for the production of ethylidene diacetate from acetic anhydride

    DOEpatents

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

    1998-01-01

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

  9. The effect of potassium lactate and sodium diacetate on the microbial, sensory, color and chemical characteristics of vacuum-packaged beef top loin steaks 

    E-print Network

    Anwar, Najia

    2000-01-01

    Beef strip loins were injected with potassium lactate (1.5, 2.0, and 2.5%), sodium diacetate (0.1%), and combination of sodium diacetate (0.1%) with 1.5 or 2.0% potassium lactate. Top loin steaks were vacuum-packaged and stored for up to 49 days...

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

    EPA Science Inventory

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

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

    EPA Science Inventory

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

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

    Microsoft Academic Search

    Jair A. Esquivel-Guzmán; Gerardo Zaragoza-Galán; Jesús Ortíz-Palacios; Ernesto Rivera

    2012-01-01

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

  13. Computer-aided modeling of the binding sites of anti-fluorescein antibodies 

    E-print Network

    Harris, Jonathan S

    1996-01-01

    SPECTROSCOPY The mouse hybridomas producing monoclonal antibodies to the fluorescein hapten conjugated through the 4' position of the xanthene ring (4'-(aminomethyl) fluorescein) to the bovine serum albumin carrier protein were previously produced... of the subclasses has their own set of constant primers used for sequencing the variable domain. To determine the IgG subclass, goat antibodies to mouse immunoglobulins conjugated to biotin were incubated with 2 ml of each hybridoma supernatant for 30 minutes...

  14. 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 ...

  15. THE EFFECT OF SODIUM LACTATE AND SODIUM DIACETATE ON THE BEHAVIOR OF LISTERIA MONOCYTOGENES IN HAM STORED AT VARIOUS TEMPERATURES

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  16. Comparison of cellulose diacetate and polysulfone membranes in the outcome of acute renal failure. A prospective randomized study

    Microsoft Academic Search

    Karine Gastaldello; Christian Melot; Robert-Jean Kahn; Jean-Louis Vanherweghem; Jean-Louis Vincent; Christian Tielemans

    survival and recovery time are not significantly influ- enced by the use of either meltspun cellulose diacetate unsubstituted cellulosic membranes is associated with a less favourable patient outcome than dialysis with of the membrane did not influence patient outcome. biocompatible synthetic membranes. Since we generally Key words: acute renal failure; biocompatibility; dia- use a modified cellulosic membrane with substantially lysis

  17. Whole Blood Cell Staining Device

    NASA Technical Reports Server (NTRS)

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

    2000-01-01

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

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

    DOEpatents

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

    1998-01-01

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

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

    DOEpatents

    Ramprasad, D.; Waller, F.J.

    1998-04-28

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

  20. Stain Free Total Protein Staining is a Superior Loading Control to ?-Actin for Western Blots

    PubMed Central

    Gilda, Jennifer E.; Gomes, Aldrin V.

    2013-01-01

    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

  1. Fluorogenic Substrate Detection of Viable Intracellular and Extracellular Pathogenic Protozoa

    NASA Astrophysics Data System (ADS)

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

    1985-01-01

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

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

    PubMed Central

    Sherr, Evelyn B.; Sherr, Barry F.

    1983-01-01

    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 PMID:16346446

  3. A comparison of fluorescent stains for the assessment of viability and metabolic activity of lactic acid bacteria.

    PubMed

    Zotta, T; Guidone, A; Tremonte, P; Parente, E; Ricciardi, A

    2012-03-01

    Lactic acid bacteria (LAB) are used as starter or probiotic cultures in the food and pharmaceutical industries and, therefore, rapid and accurate methods for the detection of their viability are of practical relevance. In this study 10 LAB strains, belonging to the genera Enterococcus, Lactococcus, Leuconostoc, Lactobacillus, Streptococcus and Weissella, were subjected to heat and oxidative stresses and cell injury or death was assessed comparing different fluorescent probes (Syto 9; Propidium Iodide, PI; 4,6-diamidino-2-phenylindole, DAPI; 5,(6)-carboxyfluorescein diacetate, cFDA) to identify the stain combination which most reliably allowed the detection of live/metabolically active and dead cells. Protocols for specimen preparation and staining were optimized and a simple procedure for automated cell counts was developed using NIH ImageJ macros. Cysteine and semi-solid agar solution were efficiently used as anti-fading agent and mounting medium, respectively. The double staining cFDA-PI apparently offered the best and most versatile indication of both cell metabolic activity and membrane integrity. An excellent correlation between manual and automated cell counts for the majority of strain/stain combinations was found. This work provides a simple protocol for specimen preparation and staining based on the use of safe, easy to prepare and inexpensive reagents as compared to other methods. Additionally, the automated cell count procedure developed can be applied to several bacterial species and allows an increase in the number of experimental trials and the reproducibility and sensitivity of the analysis. PMID:22805812

  4. Salt stains from evaporating droplets

    PubMed Central

    Shahidzadeh, Noushine; Schut, Marthe F. L.; Desarnaud, Julie; Prat, Marc; Bonn, Daniel

    2015-01-01

    The study of the behavior of sessile droplets on solid substrates is not only associated with common everyday phenomena, such as the coffee stain effect, limescale deposits on our bathroom walls , but also very important in many applications such as purification of pharmaceuticals, de-icing of airplanes, inkjet printing and coating applications. In many of these processes, a phase change happens within the drop because of solvent evaporation, temperature changes or chemical reactions, which consequently lead to liquid to solid transitions in the droplets. Here we show that crystallization patterns of evaporating of water drops containing dissolved salts are different from the stains reported for evaporating colloidal suspensions. This happens because during the solvent evaporation, the salts crystallize and grow during the drying. Our results show that the patterns of the resulting salt crystal stains are mainly governed by wetting properties of the emerging crystal as well as the pathway of nucleation and growth, and are independent of the evaporation rate and thermal conductivity of the substrates. PMID:26012481

  5. Salt stains from evaporating droplets.

    PubMed

    Shahidzadeh, Noushine; Schut, Marthe F L; Desarnaud, Julie; Prat, Marc; Bonn, Daniel

    2015-01-01

    The study of the behavior of sessile droplets on solid substrates is not only associated with common everyday phenomena, such as the coffee stain effect, limescale deposits on our bathroom walls , but also very important in many applications such as purification of pharmaceuticals, de-icing of airplanes, inkjet printing and coating applications. In many of these processes, a phase change happens within the drop because of solvent evaporation, temperature changes or chemical reactions, which consequently lead to liquid to solid transitions in the droplets. Here we show that crystallization patterns of evaporating of water drops containing dissolved salts are different from the stains reported for evaporating colloidal suspensions. This happens because during the solvent evaporation, the salts crystallize and grow during the drying. Our results show that the patterns of the resulting salt crystal stains are mainly governed by wetting properties of the emerging crystal as well as the pathway of nucleation and growth, and are independent of the evaporation rate and thermal conductivity of the substrates. PMID:26012481

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

    PubMed

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

    1979-12-01

    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. PMID:540096

  7. Fluorescein-conjugated lysine monomers for solid phase synthesis of fluorescent peptides and PNA oligomers.

    PubMed

    Lohse, J; Nielsen, P E; Harrit, N; Dahl, O

    1997-01-01

    Fluorescein ethyl ester, I, was used to prepare the fluorescent mixed ester/ether 6-O-(carboxymethyl)-fluorescein ethyl ester, III. Conjugation of III to the epsilon-amino group of alpha-N-Boc-L-lysine, via the N-hydroxysuccinimde ester, IV, gave the Boc-protected fluorescein-conjugated lysine monomer V. Removal of the Boc group, followed by reaction with Fmoc chloride, gave the Fmoc-protected monomer, VI (Figure 1). These Boc- and Fmoc-protected fluorescein-conjugated lysines were readily incorporated into peptides and PNA oligomers during solid phase synthesis to give fluorescent products. Mass spectroscopy and UV studies showed that the fluorophore remains unchanged during solid phase synthesis. In contrast to fluorescein, the photophysical properties of these derivatives are pH independent from pH 3 to 8, with a molar absorption coefficient, epsilon max 456, of 2.9 x 10(4) M-1 cm-1 and fluorescence quantum yield, phi f, of 0.18. PMID:9258448

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

    PubMed

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

    2012-08-10

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

  9. Specific immunofluorescent staining of pathogenic treponemes with a monoclonal antibody.

    PubMed Central

    Ito, F; Hunter, E F; George, R W; Pope, V; Larsen, S A

    1992-01-01

    Two hybrid cell lines which produced mouse monoclonal antibody to the DAL-1 street strain of Treponema pallidum subsp. pallidum were established. These monoclonal antibodies strongly reacted with T. pallidum subsp. pallidum (Nichols strain, DAL-1, and two other street strains, strains MN-1 and MN-3) and T. pallidum subsp. pertenue by indirect microimmunofluorescent antibody and enzyme-linked immunosorbent assay techniques, but they did not react with normal rabbit testicular tissue. These monoclonal antibodies did not react with nonpathogenic treponemes, such as T. phagedenis Reiter, T. denticola MRB, T. refringens Noguchi, or other spirochetes, such as Borrelia burgdorferi and Leptospira interrogans serovar pomona in microimmunofluorescent antibody smear slides or in Western blots (immunoblots). While unlabeled antibodies are useful for investigating the antigenic structures of T. pallidum, we labeled these monoclonal antibodies with fluorescein isothiocyanate and used them for diagnosing syphilis by direct staining of lesion exudate or T. pallidum subsp. pallidum in formalin-fixed tissues from patients suspected of having syphilis. Both monoclonal antibodies were directed against antigens of T. pallidum subsp. pallidum with a molecular weight of 37,000 as determined by the Western blotting technique. Images PMID:1374079

  10. Fluorescence spectrum analysis using Fourier series modeling for Fluorescein solution in Ethanol

    E-print Network

    Hadi, Mahasin F

    2011-01-01

    We have measured the fluorescence spectrum for fluorescein solution in ethanol with concentration 1 {\\times} 10-3 mol/liter at different temperatures from room temperature to freezing point of solvent, (T = 153, 183, 223, 253, and 303 K) using liquid nitrogen. Table curve 2D version 5.01 program has been used to determine the fitting curve and fitting equation for each fluorescence spectrum. Fourier series (3 {\\times} 2) was the most suitable fitting equation for all spectra. Theoretical fluorescence spectrum of fluorescein in ethanol at T = 183K was calculated and compared with experimental fluorescence spectrum at the same temperature. There is a good similarity between them.

  11. Problem Solving: Pencil Box Staining

    NSDL National Science Digital Library

    WGHB Boston

    2013-01-01

    This professional development video clip shows students engaged in the first Common Core Practice Standard—Make 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.

  12. A Quantitative and Standardized Method for the Evaluation of Choroidal Neovascularization Using MICRON III Fluorescein Angiograms in Rats

    PubMed Central

    Wigg, Jonathan P.; Zhang, Hong; Yang, Dong

    2015-01-01

    Introduction In-vivo imaging of choroidal neovascularization (CNV) has been increasingly recognized as a valuable tool in the investigation of age-related macular degeneration (AMD) in both clinical and basic research applications. Arguably the most widely utilised model replicating AMD is laser generated CNV by rupture of Bruch’s membrane in rodents. Heretofore CNV evaluation via in-vivo imaging techniques has been hamstrung by a lack of appropriate rodent fundus camera and a non-standardised analysis method. The aim of this study was to establish a simple, quantifiable method of fluorescein fundus angiogram (FFA) image analysis for CNV lesions. Methods Laser was applied to 32 Brown Norway Rats; FFA images were taken using a rodent specific fundus camera (Micron III, Phoenix Laboratories) over 3 weeks and compared to conventional ex-vivo CNV assessment. FFA images acquired with fluorescein administered by intraperitoneal injection and intravenous injection were compared and shown to greatly influence lesion properties. Utilising commonly used software packages, FFA images were assessed for CNV and chorioretinal burns lesion area by manually outlining the maximum border of each lesion and normalising against the optic nerve head. Net fluorescence above background and derived value of area corrected lesion intensity were calculated. Results CNV lesions of rats treated with anti-VEGF antibody were significantly smaller in normalised lesion area (p<0.001) and fluorescent intensity (p<0.001) than the PBS treated control two weeks post laser. The calculated area corrected lesion intensity was significantly smaller (p<0.001) in anti-VEGF treated animals at 2 and 3 weeks post laser. The results obtained using FFA correlated with, and were confirmed by conventional lesion area measurements from isolectin stained choroidal flatmounts, where lesions of anti-VEGF treated rats were significantly smaller at 2 weeks (p = 0.049) and 3 weeks (p<0.001) post laser. Conclusion The presented method of in-vivo FFA quantification of CNV, including acquisition variable corrections, using the Micron III system and common use software establishes a reliable method for detecting and quantifying CNV enabling longitudinal studies and represents an important alternative to conventional CNV quantification methods. PMID:26024231

  13. Methods for chromosome-specific staining

    DOEpatents

    Gray, J.W.; Pinkel, D.

    1995-09-05

    Methods and compositions for chromosome-specific staining are provided. Compositions comprise heterogeneous mixtures of labeled nucleic acid fragments having substantially complementary base sequences to unique sequence regions of the chromosomal DNA for which their associated staining reagent is specific. Methods include ways for making the chromosome-specific staining compositions of the invention, and methods for applying the staining compositions to chromosomes. 3 figs.

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

    NASA Astrophysics Data System (ADS)

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

    2013-06-01

    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.

  15. Encapsulation of T4 bacteriophage in electrospun poly(ethylene oxide)/cellulose diacetate fibers.

    PubMed

    Korehei, Reza; Kadla, John F

    2014-01-16

    Phage therapy is a potentially beneficial approach to food preservation and storage. Sustained delivery of bacteriophage can prevent bacterial growth on contaminated food surfaces. Using coaxial electrospinning bacteriophage can be encapsulated in electrospun fibers with high viability. The resulting bio-based electrospun fibers may have potential as a food packaging material. In the present work, T4 bacteriophage (T4 phage) was incorporated into core/shell electrospun fibers made from poly(ethylene oxide) (PEO), cellulose diacetate (CDA), and their blends. Fibers prepared using PEO as the shell polymer showed an immediate burst release of T4 phage upon submersion in buffer. The blending of CDA with PEO significantly decreased the rate of phage release, with no released T4 phage being detected from the solely CDA fibers. Increasing the PEO molecular weight increased the electrospun fiber diameter and viscosity of the releasing medium, which resulted in a relatively slower T4 phage release profile. SEM analyses of the electrospun fiber morphologies were in good agreement with the T4 phage release profiles. Depending on the PEO/CDA ratio, the post-release electrospun fiber morphologies varied from discontinuous fibers to minimally swollen fibers. From these results it is suggested that the T4 phage release mechanism is through solvent activation/polymer dissolution in the case of the PEO fibers and/or by diffusion control from the PEO/CDA blend fibers. PMID:24188849

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

    NASA Astrophysics Data System (ADS)

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

    2014-07-01

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

  17. Original article Blue-stain fungi associated

    E-print Network

    Paris-Sud XI, Université de

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

  18. Rapid, one step staining procedures for analysis of cellular DNA and protein by single and dual laser flow cytometry

    SciTech Connect

    Crissman, H.A.; Steinkamp, J.A.

    1982-01-01

    Detailed, simplified techniques are described for simultaneous staining and analysis of DNA and protein in a number of mammalian cell types. Cell staining involves the addition of appropriate dye mixtures to unfixed or ethanol-fixed cells and subsequent analysis of cell populations in the staining reagents generally within 10 to 20 minutes. The approach is novel in that no centrifugation steps are involved during the staining procedure, thus, eliminating cell clumping and cell loss and making the procedures appropriate for samples containing limited numbers of cells. For single wavelength analysis, staining of DNA and protein in ethanol-fixed cells was accomplished with a dye solution containing propidium iodide, fluorescein isothimocyanate and RNase. After 20 minutes at room temperature cells were analyzed using the 488 nanometer (nm) laser excitation line. For dual laser analysis the following dye combinations were employed without RNase: mithramycin-rhodamine 640, mithramycin-substituted rhodamine isothiocyanate, Hoechst 33342-rhodamine 640 and Hoechst 33342-rhodamine isothiocyanate. Unfixed cells were also stained with the Hoechst 33342-rhodamine 640 dye combination. Mithramycin was excited at 457.9 nm, Hoechst 33342 at 333-363 nm, and rhodamine dyes at 568 nm. Cell types analyzed included Chinese hamster ovary cells, cultured mouse colon 26 cells, mouse embryo forelimb bud cells, and rat cell obtained by lung lavage.

  19. Direct Nitric Oxide Detection in Aqueous Solution by Copper(II) Fluorescein Complexes

    E-print Network

    Baik, Mu-Hyun

    Direct Nitric Oxide Detection in Aqueous Solution by Copper(II) Fluorescein Complexes Mi Hee Lim nitrogen atoms of the FLn ligands most likely bind to Cu(II). Introduction of nitric oxide (NO) to pH 7-time detection of nitric oxide production in living cells. Introduction Nitric oxide (NO) is produced

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

    Microsoft Academic Search

    S S Hayreh; T A Weingeist

    1980-01-01

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

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

    PubMed

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

    2015-02-21

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

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

    PubMed Central

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

    2011-01-01

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

  3. SELECTIVE "STAINING" FOR ELECTRON MICROGRAPHY

    PubMed Central

    Mudd, Stuart; Anderson, Thomas F.

    1942-01-01

    The physical basis of contrast and image formation in electron micrography is considered in relation to the possibility of recording selective chemical effects on cell components. A technology of selective microchemical analysis, equivalent to differential staining, is suggested as practicable in electron micrography. Electron pictures of bacteria after exposure to salts of heavy metals have shown the bacterial inner protoplasm, but not the cell walls, to be selectively darkened; shrinkage, coagulation, or escape of protoplasm from the injured cells may result and be recorded in the electron micrographs. Recording of the action of germicidal agents on individual bacterial cells is indicated as one promising field of application of microchemical analysis with the aid of the electron microscope. PMID:19871216

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

    E-print Network

    Lebendiker, Mario

    SYPRO Ruby Protein Gel Stain Advanced staining technology for 2-D gels and proteomics ® Detection steps Broad linear quantitation range and consistent gel-to-gel staining allow for accurate protein especially for the analysis of proteins in 2-D polyacrylamide gels, SYPRO Ruby protein gel stain is ideal

  5. Combined silver Perls's stain for differential staining of ringed sideroblasts and marrow iron

    Microsoft Academic Search

    K T Tham; J B Cousar

    1993-01-01

    During a study of nucleolar organiser regions, a modified silver stain was found to be a sensitive marker for the iron in ringed sideroblasts, more so than Perls's stain when the marrow iron stores were low. To enhance the usefulness of the silver stain, a combined silver Perls method was developed. This stains the ringed sideroblast iron black and haemosiderin

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed

    Ellis, E Ann

    2014-01-01

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

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

    EPA Science Inventory

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

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

    EPA Science Inventory

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

  10. Effect of silver NPs plasmon on optical properties of fluorescein dye

    NASA Astrophysics Data System (ADS)

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

    2013-11-01

    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.

  11. Detection of the halogenating activity of heme peroxidases in leukocytes by aminophenyl fluorescein.

    PubMed

    Flemmig, J; Remmler, J; Zschaler, J; Arnhold, J

    2015-06-01

    The formation of hypochlorous and hypobromous acids by heme peroxidases is a key property of certain immune cells. These products are not only involved in defense against pathogenic microorganisms and in regulation of inflammatory processes, but contribute also to tissue damage in certain pathologies. After a short introduction about experimental approaches for the assessment of the halogenating activity in vitro and in cell suspensions, we are focusing on novel applications of fluorescent dye systems to detect the formation of hypochlorous acid (HOCl) in leukocytes. Special attention is directed to properties and applications of the non-fluorescent dye aminophenyl fluorescein that is converted by HOCl, HOBr, and other strong oxidants to fluorescein. This dye allows the detection of the halogenating activity in samples containing free myeloperoxidase and eosinophil peroxidase as well as in intact granulocytes using fluorescence spectroscopy and flow cytometry, respectively. PMID:25536418

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

    NASA Astrophysics Data System (ADS)

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

    2014-03-01

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

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

    PubMed

    Zhang, Li; Zhang, Xianhong

    2014-12-10

    A novel fluorescein derivative 3-bromo-5-methylsalicylaldehyde fluorescein hydrazone (BMSFH) has been synthesized by reacting fluorescein hydrazide with 3-bromo-5-methylsalicylaldehyde and was developed as a new colorimetric probe for detection of Cu(2+). In the presence of Cu(2+) the BMSFH exhibits a rapid color change from colorless to yellow together with an obvious new band appeared at 502nm in the UV-vis absorption spectra. However, other common alkali-, alkaline earth-, transition- and rare earth metal ions induced no or minimal spectral changes. This change is attributed to BMSFH via coordination with Cu(2+) in a 1:1 stoichiometry and this binding to Cu(2+) is reversible, as indicated by the bleaching of the color when the Cu(2+) is extracted with EDTA. Experimental results indicate that the BMSFH can provide a rapid, selective and sensitive response to Cu(2+) with a linear dynamic range 3.0-330?mol/L and can be used as a potential Cu(2+) colorimetric probe in aqueous solution. PMID:24929315

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

    NASA Astrophysics Data System (ADS)

    Zhang, Li; Zhang, Xianhong

    2014-12-01

    A novel fluorescein derivative 3-bromo-5-methylsalicylaldehyde fluorescein hydrazone (BMSFH) has been synthesized by reacting fluorescein hydrazide with 3-bromo-5-methylsalicylaldehyde and was developed as a new colorimetric probe for detection of Cu2+. In the presence of Cu2+ the BMSFH exhibits a rapid color change from colorless to yellow together with an obvious new band appeared at 502 nm in the UV-vis absorption spectra. However, other common alkali-, alkaline earth-, transition- and rare earth metal ions induced no or minimal spectral changes. This change is attributed to BMSFH via coordination with Cu2+ in a 1:1 stoichiometry and this binding to Cu2+ is reversible, as indicated by the bleaching of the color when the Cu2+ is extracted with EDTA. Experimental results indicate that the BMSFH can provide a rapid, selective and sensitive response to Cu2+ with a linear dynamic range 3.0-330 ?mol/L and can be used as a potential Cu2+ colorimetric probe in aqueous solution.

  15. Applications of SYPRO Orange and SYPRO Red Protein Gel Stains

    Microsoft Academic Search

    Thomas H. Steinberg; Richard P. Haugland; Victoria L. Singer

    1996-01-01

    We have further characterized the sensitivity and specificity of SYPRO Orange protein gel stain and SYPRO Red protein gel stain with native and 2-dimensional polyacrylamide gels and for staining gels prior to Western blot analysis. We found that nucleic acids are not stained by the SYPRO protein gel stains, in contrast to results obtained with commonly used silver staining techniques.

  16. Pro-Q Diamond Phosphoprotein Gel Stain

    E-print Network

    Lebendiker, Mario

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

  17. Mineral Stains at the No Name Prospect

    USGS Multimedia Gallery

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

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

    EPA Science Inventory

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

  19. Fluorescein angiography

    MedlinePLUS

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

  20. Simultaneous indocyanine green and fluorescein angiography in retinal pigment epithelium tear using the confocal scanning laser ophthalmoscope

    Microsoft Academic Search

    Ruth Axer-Siegel; Henia Lichter; Irit Rosenblatt; Ethan Priel; Yuval Yassur; Dov Weinberger

    1999-01-01

    PURPOSE:To describe the indocyanine green angiographic pattern of retinal pigment epithelium tears in the setting of age-related macular degeneration compared with the fluorescein angiographic features.METHODS:Twelve consecutive patients (12 eyes) with a retinal pigment epithelium tear underwent simultaneous indocyanine green angiography and fluorescein angiography with the confocal scanning laser ophthalmoscope. The findings for the two modes were compared.RESULTS:Choroidal neovascular membrane was

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

    PubMed

    Smith, A A; Glickfield, I

    2011-04-01

    Re-staining of formalin fixed paraffin sections sometimes is required and this requires prior de-staining. Some simple and effective protocols for de-staining are described. Mucihematoxylin and mucicarmine can be removed with acid alcohol. Zirconyl hematoxylin can be removed with periodic acid or Sinha's fixative. Alcian blue can be removed with 5% trifluoroacetic acid in dichloromethane. Colloidal iron can be bleached in 1% household bleach in alcohol. PAS can be removed with hydrogen peroxide or ammonium hydroxide. With few exceptions, de-stained sections can be re-stained with mucihematoxylin, PAS or Gabe's trichrome. PMID:20001228

  2. Lectin staining of cultured CNS microglia.

    PubMed

    Colton, C A; Abel, C; Patchett, J; Keri, J; Yao, J

    1992-04-01

    Carbohydrate binding proteins, known as lectins, bind to specific sugar groups on most membranes. We used fluorescent and light microscopy to study the interaction of various lectins with the membranes of microglia cultured from neonatal rat or fetal mouse cerebral cortices. Microglia stained intensely with GS-1, RCA, WGA, and ConA and slightly with DBA, UEA, BPA, and SBA. No staining was seen with GS-2, MPA, or PNA. Staining was specific for microglia in the mixed glial cultures and was dose dependent. In addition, microglial lectin binding could be reduced or blocked by competitive inhibition using specific sugars. Treatment of the microglia with agents such as dimethylsulfoxide (DMSO), interleukin-1 (IL-1), interferon (IFN), or lipopolysaccharide (LPS) did not eliminate lectin staining, although the degree of staining was altered. Positive staining of the microglia was also associated with a functional change for at least one lectin, i.e., ConA. Superoxide anion production by microglia was increased in the presence of ConA. Overall, binding of the lectins GS-1, RCA, WGA, and ConA can be used as an identifying tool for microglia in glial cultures, but intensity of staining varies depending on their functional state. PMID:1372634

  3. Comparison of Special Stains for Keratin with Routine Hematoxylin and Eosin Stain

    PubMed Central

    Rao, Roopa S; Patil, Shankargouda; Majumdar, Barnali; Oswal, Rakesh G

    2015-01-01

    Background: Keratins are the most abundant proteins and are characteristic findings in many epithelial pathologies, making it a diagnostically important marker, both histopathologically and immunohistochemically. Since, immunohistochemistry is an expensive diagnostic tool, special stains to detect the degree of keratinization could serve as a faster and economic option. The aim of the present study was to compare the efficacy of special stains for keratin with standard hematoxylin and eosin stain (H and E). Objectives include: (i) To subject the diagnosed cases of keratin disorders to the selected special stains: Ayoub-shklar method, Dane-Herman method, Alcian blue –periodic acid Schiff ’s (PAS), rapid papanicolaou (PAP) and Gram’s stain. (ii) To compare the staining specificity and staining intensity of special stains with respect to routine hematoxylin and eosin (H and E) stain. (iii) To compare the efficacy of special stains to routine H and E stain in identification of the type of keratin present in the selected cases. Materials and Methods: A total of 80 cases of known pathology for keratin were retrieved from the department archive, which included 10 each of normal gingiva, hyperkeratosis, squamous papilloma, verrucous hyperplasia, verrucous carcinoma, well-differentiated squamous cell carcinoma, orthokeratinized odontogenic cyst and keratocystic odontogenic tumors. Six sections of 4 µ each from the paraffin blocks were made, stained with H and E and the special stains and these were evaluated by 2 pathologists based on the modified scoring criteria from Rahma Al-Maaini and Philip Bryant 2008. Results: The results were tabulated using Chi square and kappa statistics. The statistical values for identification of the type of keratinization was insignificant showing that ortho and parakeratinized epithelia could be correctly identified by both H and E as well as all the special stains. Furthermore, all the special stains showed a positive result and statistical significance (P < 0.001) with respect to the staining of keratin. Conclusion: To conclude, though the special stains distinctly stained the keratin with a higher intensity, H and E proves to be overall better stain with respect to specificity PMID:25878469

  4. An alternative to India ink stain.

    PubMed

    Ibembe, Isaac Nicholas; Wiggin, Timothy Roger

    2015-07-01

    Accessing India ink in rural Uganda is difficult and costly. An alternative stain was sought to assist in microbiological diagnoses of cryptococcal infections in immunosuppressed patients with meningitis. Mascara proved to be an excellent and cheap alternative. PMID:25999353

  5. A ‘Magnetic’ Gram Stain for Bacterial Detection

    PubMed Central

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

    2012-01-01

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

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

    PubMed Central

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

    2014-01-01

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

  7. New Grocott Stain without Using Chromic Acid

    PubMed Central

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

    2015-01-01

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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  9. 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...

  10. COMPARATIVE STUDIES OF THE STRUCTURES AND TRANSITION CHARACTERISTICS OF CELLULOSE DIACETATE MODIFIED WITH POLYETHYLENE GLYCOL PREPARED BY CHEMICAL BONDING AND PHYSICAL BLENDING METHODS

    Microsoft Academic Search

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

    2001-01-01

    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

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

    PubMed Central

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

    2013-01-01

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

  12. Tailoring of optical properties of fluorescein using green synthesized gold nanoparticles.

    PubMed

    John, Jisha; Thomas, Lincy; George, Nibu A; Kurian, Achamma; George, Sajan D

    2015-06-28

    Dye-nanoparticle mixtures hold great promise in biological as well as photonics applications due to their capability to tailor the emission behavior of dye by tuning the nanoparticles parameters. However, as compared to the well-defined dye-nanoparticle distance, studies lack the understanding of homogenous mixtures of dye and nanoparticles. In this work, we investigate the influence of shape and concentration of gold nanoparticles prepared via green synthesis on the optical properties of fluorescein dye in a dye-nanoparticle mixture. We have investigated the radiative path of deexcitation using steady state fluorescence and the non-radiative path is probed using a laser based dual-beam thermal lens technique. The energy transfer efficiency as well as dye-nanoparticle distance is studied using both techniques. Furthermore, we have explored the influence of nanoparticles parameters on the fluorescence quantum yield of fluorescein using the thermal lens technique. The studies indicate that spherical nanoparticles are efficient quenchers while star shaped nanoparticles can probe larger dye-NP distances. The tailoring of dye properties by tuning nanoparticle parameters can be utilized in diverse areas including bioimaging, solar cells, and sensors. PMID:26017461

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

    PubMed Central

    Hayreh, S S; Weingeist, T A

    1980-01-01

    Transient experimental occlusion of the central artery of the retina (OCAR), lasting from 15 to 270 minutes, was produced by clamping the artery in the orbit in 63 eyes of rhesus monkeys. Ophthalmoscopic and fluorescein angiographic studies were performed before and during clamping of the artery, as well as periodically after unclamping, for periods of up to 22 weeks. The effects of transient retinal ischaemia on the retina, optic disc, and retinal vascular bed were studied. 89% of the eyes showed a variable amount of residual retinal circulation on angiography during CAR clamping, but this did not exercise any protective action against ischaemic damage. Duration of the ischaemia was the principal factor determining severity of damage. OCAR for up to 98 minutes produced no significant permanent neural damage, but OCAR for 105 minutes or longer produced irreversible permanent neural damage. There was no significant permanent damage to the retinal vascular bed, though a transient fluorescein leakage was seen after OCAR for 2 1/2-3 hours or longer. The findings revealed that the normal red colour of the optic disc represents retinal vascular filling in the surface layer of the disc and not deeper vascular filling. The various factors influencing the retinal circulation and neural damage in OCAR are discussed. Images PMID:7448143

  14. Physiological and pathobiological significance of ocular glycoproteins. I. Studies using fluorescein labelled glycine max.

    PubMed

    Ahmed, A I; Rahi, A H

    1985-03-01

    Cell surface carbohydrates play an important role in several biological, immunological, and neoplastic phenomena including development, growth regulation, cellular locomotion, receptor activation, and tumour metastasis. Fluorescein labelled lectins which bind to specific carbohydrate residues in glycoproteins and glycolipids are being increasingly used as chemical probes to study cell components. Several different preparations of ocular tissues from human, rabbit, and rat were examined for the distribution of N-acetyl-D-galactosamine (D-gal NAc) by means of fluorescein-labelled lectin from soybean (glycine max). A very strong fluorescence was observed in the corneal epithelium; Descemet's membrane and corneal endothelium were also strongly fluorescent. The conjunctival epithelium similarly showed a strong reaction, as did the goblet cells. The iris epithelium and the dilator pupillae were only weakly fluorescent, but the ciliary body showed strong fluorescence, as did the blood vessels. As compared with lens fibres the lens epithelium was strongly fluorescent. The outer retina, that is, the photoreceptors, the pigment epithelium, and Bruch's membrane, showed a very strong reactivity. The optic nerve showed moderate fluorescence, but reaction with extraocular muscles was variable. The skin of the upper and lower eyelids, hair follicles, and blood vessels showed strong lectin binding. Sections of retinoblastoma and malignant melanoma showed no reaction. The physiological and pathological significance of these findings is discussed. PMID:4038883

  15. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

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

  16. Soluble oligovalent antigen-antibody complexes. I. The effect of antigen valence and combining ratio on the composition of fluorescein-carrier anti-fluorescein complexes.

    PubMed Central

    van Es, L A; Knutson, D W; Kayser, B S; Glassock, R J

    1979-01-01

    Soluble oligovalent antigen--antibody complexes were prepared and analysed by ultracentrifugation in order to study the effect of the combining ratio, antigen valence and concentration upon the size and molecular composition of the composition of the complexes. Fluorescein (F) conjugates of rabbit serum albumin (RSA) and thyroglobulin (RTg) were combined with high affinity rabbit anti-F antibodies to form soluble complexes. The effect of the combining ratio paralleled findings in precipitating systems in that the largest soluble complexes were found at equimolarity and mild molar antibody excess. Tetravalent antigen formed precipitates at combining ratios near equimolarity, whereas trivalent antigens failed to precipitate at similar concentrations. Complexes prepared near equimolarity were most sensitive to changes in concentration, higher concentrations leading to larger complexes. The Ab/Ag ratios of different-size complexes in the same preparation were remarkably similar. This ratio was dependent on the antibody--antigen combining ratio, was limited by antigen valence and was not affected by concentration differences. The data support the hypothesis that soluble complexes are formed in two steps. First, antigen and antibody combine to form subunits whose Ab/Ag ratio is determined by the combining ratio and antigen valence. These subunits then combine to form larger complexes in a manner analogous to polymerization. PMID:468312

  17. Compositions for chromosome-specific staining

    DOEpatents

    Gray, J.W.; Pinkel, D.

    1998-05-26

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. The methods produce staining patterns that can be tailored for specific cytogenetic analyses. The probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. The invention provides for automated means to detect and analyze chromosomal abnormalities. 17 figs.

  18. Compositions for chromosome-specific staining

    DOEpatents

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

    1998-01-01

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Further, methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. Still further, the invention provides for automated means to detect and analyse chromosomal abnormalities.

  19. Artifactual Sulfation of Silver-stained Proteins

    PubMed Central

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

    2009-01-01

    Sulfation and phosphorylation are post-translational modifications imparting an isobaric 80-Da addition on the side chain of serine, threonine, or tyrosine residues. These two post-translational modifications are often difficult to distinguish because of their similar MS fragmentation patterns. Targeted MS identification of these modifications in specific proteins commonly relies on their prior separation using gel electrophoresis and silver staining. In the present investigation, we report a potential pitfall in the interpretation of these modifications from silver-stained gels due to artifactual sulfation of serine, threonine, and tyrosine residues by sodium thiosulfate, a commonly used reagent that catalyzes the formation of metallic silver deposits onto proteins. Detailed MS analyses of gel-separated protein standards and Escherichia coli cell extracts indicated that several serine, threonine, and tyrosine residues were sulfated using silver staining protocols but not following Coomassie Blue staining. Sodium thiosulfate was identified as the reagent leading to this unexpected side reaction, and the degree of sulfation was correlated with increasing concentrations of thiosulfate up to 0.02%, which is typically used for silver staining. The significance of this artifact is discussed in the broader context of sulfation and phosphorylation site identification from in vivo and in vitro experiments. PMID:18936056

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

    E-print Network

    Boyer, Edmond

    IN VIVO STAINING OF ASTROCYTES 1 Specific in vivo staining of astrocytes in the whole brain after of astrocytes without damaging or interfering with normal brain functions is essential for intravital microscopy in vivo studies of astrocytes in the intact brain [10,11,12] and has opened a new field of dynamic

  1. Negative Stain Electron Microscopy of Microtubules Negative staining is a rapid, qualitative method for analyzing

    E-print Network

    Mitchison, Tim

    . Filtered stain stored at 4¡C in a foil-wrapped tube can be used for >1 year.) Filter strips (prepared by cutting Whatman #1 filter paper into small slivers) Grids (200 mesh copper grids that have been formvar coated, carbon coated) Rinse (ddH2O with 5 mM EGTA or as appropriate) II. Negative Staining Protocol 1

  2. Laser treatment of port-wine stains

    PubMed Central

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

    2015-01-01

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

  3. The composition of Erythrosins, Fluorescein, Phloxine and Rose Bengal: a study using thin-layer chromatography and solvent extraction

    Microsoft Academic Search

    P. N. Marshall

    1976-01-01

    Synopsis  Commercial samples of Erythrosin B (CI 45430), Erythrosin Y (CI 45425), Fluorescein (CI 45350), Phloxine (CI 45410) and Rose Bengal (CI 45440) have been analysed by thin-layer chromatography. The Erythrosins were found to be mixtures consisting in the main of 4-iodofluorescein, 4,5-di-iodofluorescein, 2,4,5-triiodofluorescein and 2,4,5,7-tetraiodofluorescein, in some instances together with 2,4,5-tri-iodo-4,5,6,7-tetrachlorofluorescein and 2,4,5,7-tetraiodo-4,5,6,7-tetrachlorofluorescein. Samples of Fluorescein were mixtures of the

  4. Meconium Staining and Meconium Aspiration Syndrome

    Microsoft Academic Search

    Sean C. Blackwell; Carlos A. Carreno; Sonia S. Hassan; Julie Moldenhauer; Honor M. Wolfe; Stanley M. Berry; Michael Kruger; Yoram Sorokin

    2001-01-01

    Objective: To determine whether the incidence of pregnancies complicated by meconium-stained amniotic fluid (MSAF) or meconium aspiration syndrome (MAS) differs with seasonal changes. Methods: An established perinatal database was used to identify all term (?37 weeks) singleton gestations resulting in a live birth from January 1, 1997 to December 31, 1999. Patients were divided into groups based on the season

  5. Crystalline lens capsule staining with trypan blue

    Microsoft Academic Search

    Joseph San Laureano; Minas T. Coroneo

    2004-01-01

    A 1-step method for staining the anterior lens capsule with trypan blue is described. The dye is instilled via a paracentesis port at the start of cataract extraction. As aqueous humor is allowed to exit the anterior chamber (AC), which consequently shallows, the resulting pupil block confines the dye to the AC. An ophthalmic viscosurgical device (OVD) is used to

  6. Automatic counting of immunocytochemically stained cells

    Microsoft Academic Search

    F. Arambula Cosio; J. A. Marquez Flores; M. A. Padilla Castaneda; S. Solano; P. Tato

    2003-01-01

    In this work is described the development of an automatic color image segmentation and cell counting system for immunocytochemical analysis of stained tissue samples. The system is designed to automatically count the total number of positive and negative cells in tissue samples treated with cytokines DNA probes of pigs naturally parasitized with Taenia solium metacestodes and using in situ hybridization.

  7. Staining and Imaging an Agarose Gel

    NSDL National Science Digital Library

    Hinkley, Craig

    This video from CUNY Kingsborough Community College shows the process of staining and imaging an agarose gel. The video describes the process step by step and would be easy to replicate in a laboratory setting. Running time for the video is 3:18.

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

    PubMed

    Meyers, B; Valencia, S

    1989-01-01

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

  9. Improving vessel segmentation in ultra-wide field-of-view retinal fluorescein angiograms.

    PubMed

    Perez-Rovira, A; Zutis, K; Hubschman, J P; Trucco, E

    2011-01-01

    Vessel segmentation on ultra-wide field-of-view fluorescein angiogram sequences of the retina is a challenging problem. Vessel appearance undergoes severe changes, as different portions of the vascular structure become perfused in different frames. This paper presents a method for segmenting vessels in such sequences using steerable filters and automatic thresholding. We introduce a penalization stage on regions with high vessel response in the filtered image, improving the detection of peripheral vessels and reducing false positives around the optic disc and in regions of choroidal vessels and lesions. Quantitative results are provided, in which the penalization stage improves the segmentation precision segmentation by 11.84%, the recall by 12.98% and the accuracy by 0.40%. To facilitate further evaluation, usage, and algorithm comparison, the algorithm, the data set used, the ground truth, and the results are made available on the internet. PMID:22254877

  10. Computer-aided modeling of the binding sites of anti-fluorescein antibodies

    E-print Network

    Harris, Jonathan S

    1996-01-01

    V the interactions between the antibody and the fluorescein molecule, Binding of an antibody to antigen mu'st overcome the negative effect from entropic contributions. This can occur by formation of hydrogen bonds, van der Waals' contacts... CI CI LU 0 ID N IU O 0 CI O OO 0 '0 UO C al od C IO V 0 al 0 '0 O al ) 0 C 0 aa I V V V 0 II al Cd cd 'a al 'a 0 0 dl 0 0 I 0 O 0 0 0 0 al '0 al 2 V '0 0 V UO 0 OO 0 0 0 CP U V + I/I OO + FO...

  11. Automated Detection of Leakage in Fluorescein Angiography Images with Application to Malarial Retinopathy

    PubMed Central

    Zhao, Yitian; J. C. MacCormick, Ian; G. Parry, David; Leach, Sophie; A. V. Beare, Nicholas; P. Harding, Simon; Zheng, Yalin

    2015-01-01

    The detection and assessment of leakage in retinal fluorescein angiogram images is important for the management of a wide range of retinal diseases. We have developed a framework that can automatically detect three types of leakage (large focal, punctate focal, and vessel segment leakage) and validated it on images from patients with malarial retinopathy. This framework comprises three steps: vessel segmentation, saliency feature generation and leakage detection. We tested the effectiveness of this framework by applying it to images from 20 patients with large focal leak, 10 patients with punctate focal leak, and 5,846 vessel segments from 10 patients with vessel leakage. The sensitivity in detecting large focal, punctate focal and vessel segment leakage are 95%, 82% and 81%, respectively, when compared to manual annotation by expert human observers. Our framework has the potential to become a powerful new tool for studying malarial retinopathy, and other conditions involving retinal leakage. PMID:26030010

  12. Automated detection of leakage in fluorescein angiography images with application to malarial retinopathy.

    PubMed

    Zhao, Yitian; J C MacCormick, Ian; G Parry, David; Leach, Sophie; A V Beare, Nicholas; P Harding, Simon; Zheng, Yalin

    2015-01-01

    The detection and assessment of leakage in retinal fluorescein angiogram images is important for the management of a wide range of retinal diseases. We have developed a framework that can automatically detect three types of leakage (large focal, punctate focal, and vessel segment leakage) and validated it on images from patients with malarial retinopathy. This framework comprises three steps: vessel segmentation, saliency feature generation and leakage detection. We tested the effectiveness of this framework by applying it to images from 20 patients with large focal leak, 10 patients with punctate focal leak, and 5,846 vessel segments from 10 patients with vessel leakage. The sensitivity in detecting large focal, punctate focal and vessel segment leakage are 95%, 82% and 81%, respectively, when compared to manual annotation by expert human observers. Our framework has the potential to become a powerful new tool for studying malarial retinopathy, and other conditions involving retinal leakage. PMID:26030010

  13. Validation of an automated fluorescein method for determining bromide in water

    USGS Publications Warehouse

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

    1985-01-01

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

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

    PubMed Central

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

    2013-01-01

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

  15. Improved method for combination of immunocytochemistry and Nissl staining

    Microsoft Academic Search

    Andrea Kádár; Gábor Wittmann; Zsolt Liposits; Csaba Fekete

    2009-01-01

    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

  16. Separation of polyethylene glycols and their fluorescein-labeled compounds depending on the hydrophobic interaction by high-performance liquid chromatography

    Microsoft Academic Search

    Min Liu; Cao Xie; Hong Pan; Jun Pan; Weiyue Lu

    2006-01-01

    The separation and characterization of fluorescein-labeled polyethylene glycols (PEG) is described. Firstly, the polyethylene glycols labeled with fluorescein isothiocyanate (FITC) were synthesized and separated using Sephadex™ LH-20 medium by a step gradient. Secondly, a TSK GEL G4000 PWXL column was developed for determining the FITC derivatives of PEG. The retention mechanism is based on the hydrophobic interaction between the FITC

  17. SeeBand Protein Staining solution Handbook www.geba.org 1 SeeBand Protein Staining solution

    E-print Network

    Lebendiker, Mario

    staining solution guarantees protein staining in polyacrylamide gels with only slight fixation of the protein to the gel. This unique staining solution allows electro-elution of a desired protein from solution vigorously. Sensitivity: 38 ng protein per band. Gel staining procedure: Note: If fixation

  18. Improved Whole-Blood-Staining Device

    NASA Technical Reports Server (NTRS)

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

    2012-01-01

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

  19. Competition-based cellular peptide binding assays for 13 prevalent HLA class I alleles using fluorescein-labeled synthetic peptides

    Microsoft Academic Search

    Jan H Kessler; Bregje Mommaas; Tuna Mutis; Ivo Huijbers; Debby Vissers; Willemien E Benckhuijsen; Geziena M. Th Schreuder; Rienk Offringa; Els Goulmy; Cornelis J. M Melief; Sjoerd H van der Burg; Jan W Drijfhout

    2003-01-01

    We report the development, validation, and application of competition-based peptide binding assays for 13 prevalent human leukocyte antigen (HLA) class I alleles. The assays are based on peptide binding to HLA molecules on living cells carrying the particular allele. Competition for binding between the test peptide of interest and a fluorescein-labeled HLA class I binding peptide is used as read

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

    PubMed

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

    2013-08-01

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

  1. Effect of alumina on triethylene glycol diacetate-2-propenoic acid butyl ester composite polymer electrolytes for flexible lithium ion batteries

    NASA Astrophysics Data System (ADS)

    Wang, Qiujun; Song, Wei-Li; Fan, Li-Zhen; Shi, Qiao

    2015-04-01

    Triethylene glycol diacetate-2-propenoic acid butyl ester (TEGDA-BA) based composite polymer electrolytes (CPE) are fabricated by incorporating alumina (Al2O3) nanoparticles (average particle size 10-20 nm) as inorganic filler via in situ polymerization. Effects of Al2O3 concentration on ionic conductivities, Li+ transfer numbers and charge/discharge properties are studied in details. Due to the uniformly dispersed Al2O3 nanoparticles, significant improvements in the mechanical flexibility and bendability are presented in the resulting polymer electrolytes. The CPE with 5 wt% Al2O3 nanoparticles exhibits the highest ionic conductivity up to 6.02 × 10-3 S cm-1 at 25 °C and the highest Li+ transference number (0.675), coupled with the most stable electrochemical window (>4.5 V vs. Li/Li+). With the presence of Al2O3, the growth of interface resistance is retarded, which increases the interface stability. The Li|CPE|Li4Ti5O12 and Li|CPE|LiFePO4 cells demonstrate remarkably stable charge/discharge performance and excellent capacity retention during cycling test. The results suggest that the CPE holds great application potential in flexible lithium ion batteries.

  2. Automatic analysis of immunocytochemically stained tissue samples

    Microsoft Academic Search

    Fernando Arámbula Cosío; J. A. Márquez Flores; Miguel A. Padilla Castañeda; S. Solano; P. Tato

    2005-01-01

    An automatic colour image segmentation and cell counting software system has been developed for immunocytochemical analysis\\u000a of stained tissue samples. The system was designed to count the total number of positive and negative cells in tissue samples\\u000a treated with cytokine DNA probes from pigs naturally parasitised with Taenia solium metacestodes, using in situ hybridisation.\\u000a A reaction index was calculated as

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

    NASA Technical Reports Server (NTRS)

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

    1989-01-01

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

  4. Capillary electrophoresis/laser-induced fluorescence detection of fluorescein as a groundwater migration tracer.

    PubMed

    Ferguson, P L; Grange, A H; Brumley, W C; Donnelly, J R; Farley, J W

    1998-09-01

    Capillary electrophoresis (CE) has been applied to the determination of the groundwater migration tracer dye fluorescein based on laser-induced fluorescence (LIF) detection and compared to determinations obtained with traditional spectrofluorimetry. Detection limits of injected dye in the low parts per trillion (ppt) ranges have been accomplished with both CE/LIF based on the Ar ion laser and with a spectrofluorimeter. This approach was used for a real-world problem in determining groundwater migration between adjacent Resource Conservation and Recovery Act (RCRA) and Superfund sites by the Environmental Sciences Division in response to regional needs and as application of new analytical tools under development. Fluorescent dye was injected into source wells and then was determined in monitoring wells by extracting pads that adsorbed the dye or by directly determining the dye in the water using solid-phase extraction (SPE), a preconcentration technique. The approaches based on CE/LIF exhibits increased specificity over existing approaches due to the separation and unique migration time of the dye. Additional studies were aimed at achieving sub-ppt levels in the water using solid-phase extraction and field-amplified injection techniques. PMID:9761212

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

    PubMed Central

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

    2014-01-01

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

  6. Automated Detection of Vessel Abnormalities on Fluorescein Angiogram in Malarial Retinopathy

    PubMed Central

    Zhao, Yitian; MacCormick, Ian J. C.; Parry, David G.; Beare, Nicholas A. V.; Harding, Simon P.; Zheng, Yalin

    2015-01-01

    The detection and assessment of intravascular filling defects is important, because they may represent a process central to cerebral malaria pathogenesis: neurovascular sequestration. We have developed and validated a framework that can automatically detect intravascular filling defects in fluorescein angiogram images. It first employs a state-of-the-art segmentation approach to extract the vessels from images and then divide them into individual segments by geometrical analysis. A feature vector based on the intensity and shape of saliency maps is generated to represent the level of abnormality of each vessel segment. An AdaBoost classifier with weighted cost coefficient is trained to classify the vessel segments into normal and abnormal categories. To demonstrate its effectiveness, we apply this framework to 6,358 vessel segments in images from 10 patients with malarial retinopathy. The test sensitivity, specificity, accuracy, and area under curve (AUC) are 74.7%, 73.5%, 74.1% and 74.2% respectively when compared to the reference standard of human expert manual annotations. This performance is comparable to the agreement that we find between human observers of intravascular filling defects. Our method will be a powerful new tool for studying malarial retinopathy. PMID:26053690

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

    PubMed

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

    2014-07-01

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

  8. Automated Detection of Vessel Abnormalities on Fluorescein Angiogram in Malarial Retinopathy.

    PubMed

    Zhao, Yitian; MacCormick, Ian J C; Parry, David G; Beare, Nicholas A V; Harding, Simon P; Zheng, Yalin

    2015-01-01

    The detection and assessment of intravascular filling defects is important, because they may represent a process central to cerebral malaria pathogenesis: neurovascular sequestration. We have developed and validated a framework that can automatically detect intravascular filling defects in fluorescein angiogram images. It first employs a state-of-the-art segmentation approach to extract the vessels from images and then divide them into individual segments by geometrical analysis. A feature vector based on the intensity and shape of saliency maps is generated to represent the level of abnormality of each vessel segment. An AdaBoost classifier with weighted cost coefficient is trained to classify the vessel segments into normal and abnormal categories. To demonstrate its effectiveness, we apply this framework to 6,358 vessel segments in images from 10 patients with malarial retinopathy. The test sensitivity, specificity, accuracy, and area under curve (AUC) are 74.7%, 73.5%, 74.1% and 74.2% respectively when compared to the reference standard of human expert manual annotations. This performance is comparable to the agreement that we find between human observers of intravascular filling defects. Our method will be a powerful new tool for studying malarial retinopathy. PMID:26053690

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

    PubMed Central

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

    2015-01-01

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

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

    SciTech Connect

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

    1980-01-01

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

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

    PubMed

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

    2005-10-01

    We used qualitative and quantitative fluorescence microscopy of the fluorescein isothiocyanate-conjugated lectins Concanavalin A, phytohaemagglutinin-erythroagglutinin, pokeweed mitogen, and peanut agglutinin to examine sugar composition on the cell surface and cell-associated mucilage (where present) in a number of cultured and environmental algae. Lectin-binding activity was markedly different between laboratory-cultured and environmental samples of the same species. Sugar composition of the cyanobacterium Anabaena cylindrica varied with growth cycle, although no clear pattern of change was observed. Akinetes typically showed lectin-binding activity higher than that of the vegetative cells or heterocysts throughout the growth cycle. Algae with mucilage showed greater lectin binding, indicating that mucilage contained more surface sugars accessible to the lectin probe compared with the cell wall surface. A low level of galactose and N-acetyl galactosamine (detected by peanut agglutinin) was associated with the surface mucilage of most algal species. Relatively high amounts of mannose, glucose, and N-acetyl glucosamine (detected by Concanavalin A, phytohaemagglutinin, and pokeweed mitogen) were also present. Lectin binding was shown to be a highly specific and sensitive approach to the examination of cell surface chemistry of both cultured and environmental algae and to the study of biodiversity in phytoplankton. PMID:16228900

  12. Inhibition of tonoplast ATPase from etiolated mung bean seedlings by fluorescein 5'-isothiocyanate.

    PubMed Central

    Tzeng, C M; Hsu, L H; Pan, R L

    1992-01-01

    Fluorescein 5'-isothiocyanate (FITC) was used to modify the lysine residue in the active site of tonoplast H(+)-ATPase from etiolated mung-bean (Vigna radiata L.) seedlings. FITC caused marked inactivation of the enzyme activities of both membrane-bound and soluble ATPase and its associated H+ translocation. The SDS/PAGE pattern revealed that the FITC-binding site was in the large (A) subunit of ATPase. Inhibition could be substantially prevented by its physiological substrate ATP, pyrophosphate and nucleotides in the decreasing order: ATP greater than pyrophosphate greater than ADP greater than AMP greater than GTP greater than CTP greater than UTP. The mode of inhibition by FITC was competitive with respect to ATP. Loss of ATPase activity followed pseudo-first-order kinetics with a Ki of 0.33 mM, a minimum inactivation half-time of 110 s, and a first-order rate constant of 0.244 s-1. A double-logarithmic plot of apparent rate constant versus FITC concentration gave a slope of 0.913, indicating that inactivation results from reaction of at least one lysine residue at the catalytic site of the large subunit. Labelling studies indicated that the incorporation of approx. 1 mol of FITC/mol of ATPase is sufficient to inhibit ATPase completely. The enhancement and blue shift of emission maxima of FITC after modification of ATPase indicated that the labelled lysine residue was located in a relatively hydrophobic domain. PMID:1386733

  13. Correlation between Fluorescein Angiographic Findings and Visual Acuity in Behçet Retinal Vasculitis

    PubMed Central

    Kim, Min; Kwon, Hee Jung; Choi, Eun Young; Kim, Sung Soo; Koh, Hyoung Jun

    2015-01-01

    Purpose To identify significant fluorescein angiographic (FA) characteristics associated with visual acuity (VA) in Behçet retinal vasculitis. Materials and Methods Retrospective review of 86 eyes of 48 patients (age: 35.6±10.2 years) with Behçet retinal vasculitis were performed. VA and FA findings as well as correlation between them were assessed. Results The mean initial VA of eyes with posterior pole-involved vasculitis (63 eyes; 73.3%) was significantly worse than that of those with peripheral vasculitis (23 eye; 26.7%) (logarithm of the minimum angle of resolution VA: 0.554±0.572 vs. 0.078±0.148; p<0.0001). Subgroup analysis revealed a more severe and diffuse pattern of vascular leakage in posterior pole-involved vasculitis compared to peripheral vasculitis (p<0.0001). Retinal vascular leakage (?=0.345; p<0.0001), optic disc hyperfluorescence (?=0.147; p=0.032), and macular leakage (?=0.107; p=0.047) were significantly associated with worse initial VA. During the follow up (mean: 33.3±17.9 months), the change of leakage showed no significant correlation with change of VA in posterior pole-involved vasculitis (?=0.199, p=0.092). Conclusion Posterior pole involvement, the degree of retinal vascular leakage, optic disc hyperfluorescence, and macular leakage are significantly associated with VA in Behçet retinal vasculitis. PMID:26069134

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

    MedlinePLUS

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

  15. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

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

  16. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

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

  17. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

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

  18. 21 CFR 864.1850 - Dye and chemical solution stains.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

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

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

    PubMed Central

    Makin, T.; Hart, C. A.

    1989-01-01

    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 PMID:2673821

  20. Discriminative staining methods for the nervous system: luxol fast blue--periodic acid-Schiff--hematoxylin triple stain and subsidiary staining methods.

    PubMed

    Goto, N

    1987-09-01

    This paper describes a new series of staining methods which can discriminatively demonstrate every structure of the nervous system, including axons and capillaries, in animal and human materials. Methods described in this paper consist of one primary stain, luxol fast blue-periodic acid Schiff-hematoxylin (LPH) and six different subsidiary staining methods. The LPH triple stain can precisely differentiate the following structures: neurons (Nissl bodies, cytoplasm, nuclear membrane and nucleolus), various kinds of nuclei (glia, ependyma, endothelium, leucocyte, connective tissue, etc.), myelin sheaths, neuronal processes (axons and dendrites), reacted glial cell bodies (protoplasmic astrocytes, foamy cells, etc.), blood vessels (arteries, veins and capillaries), meninges, intervening connective tissue, erythrocytes, lipofuscin granules, amyloid bodies, and others. Subsidiary staining methods are also described briefly. Applications are discussed in the context of staining technology and neuromorphological research. PMID:2447684

  1. Haemodialysis related osteomalacia: a staining method to demonstrate aluminium

    Microsoft Academic Search

    Malcolm RC Buchanan; Benno U Ihle; Cheryl M Dunn

    1981-01-01

    A slight modification in tissue processing and staining technique enables a previously described method for staining aluminium to be used to demonstrate aluminium in osteomalacia associated with haemodialysis. The stain appears to be accurate in diagnosing this condition and may assist in establishing the diagnosis before severe osteomalacia develops.

  2. Cigarette staining and cleaning of a maxillofacial silicone

    SciTech Connect

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

    1983-07-01

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

  3. Organization and spatial arrangement of fluorescein-labeled native actin microinjected into normal locomoting and experimentally influenced Amoeba proteus

    Microsoft Academic Search

    Wolfgang Gawlitta; Wilhelm Stockem; Jfirgen Wehland; Klaus Weber

    1980-01-01

    Fully polymerization-competent fluorescein-labeled actin from skeletal muscle was microinjected into both normal moving and experimentally treated Amoeba proteus. Its intracellular distribution was followed by integral image intensification of the fluorescence on a television screen and compared with controls injected with rhodamine-labeled serum albumin. The labeled actin was incorporated into the endogenous actin pool and exhibited a characteristic redistribution depending on

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

    Microsoft Academic Search

    Mariana Voicescu; Marilena Vasilescu; Titus Constantinescu; Aurelia Meghea

    2002-01-01

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

  5. Antioxidant capacity of herbal infusions and tea extracts: A comparison of ORAC-fluorescein and ORAC-pyrogallol red methodologies

    Microsoft Academic Search

    E. Alarcón; A. M. Campos; A. M. Edwards; E. Lissi; C. López-Alarcón

    2008-01-01

    Oxygen radical absorbance capacity (ORAC) values have been obtained for a series of teas and herbal infusions employing 2,2?-azo-bis(2-amidinopropane) as free radical source, and fluorescein and pyrogallol red as target molecules. The amounts of phenols in the extracts were evaluated by Folin’s methodology. ORAC values are extremely dependent upon the employed target molecule. Even more, relative ORAC values measured for

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

    Microsoft Academic Search

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

    2003-01-01

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

  7. Retinal vein-to-vein anastomoses in Sturge-Weber syndrome documented by ultra-widefield fluorescein angiography.

    PubMed

    Quan, Ann V; Moore, Grant H; Tsui, Irena

    2015-06-01

    We report the case of a 6-year-old boy with Sturge-Weber syndrome and unilateral glaucoma in his left eye. He was born with a port wine mark involving his upper left eyelid. On ultra-widefield fluorescein angiography, he was found to have several vein-to-vein anastomoses in his left retina. To our knowledge, this is the first documentation of retinal vein-to-vein anastomoses in Sturge-Weber syndrome. PMID:25944745

  8. The Edge-Driven Dual-Bootstrap Iterative Closest Point Algorithm for Registration of Multimodal Fluorescein Angiogram Sequence

    Microsoft Academic Search

    Chia-Ling Tsai; Chun-Yi Li; Gehua Yang; Kai-Shung Lin

    2010-01-01

    Motivated by the need for multimodal image registration in ophthalmology, this paper introduces an algorithm which is tailored to jointly align in a common reference space all the images in a complete fluorescein angiogram (FA) sequence, which contains both red-free (RF) and FA images. Our work is inspired by Generalized Dual-Bootstrap Iterative Closest Point (GDB-ICP), which rank-orders Lowe keypoint matches

  9. Staining Protocols for Human Pancreatic Islets

    PubMed Central

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

    2012-01-01

    Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and beta mass ranges from 500 to 1500 mg 1-3. With this known heterogeneity, a standard processing and staining procedure was developed so that pancreatic regions were clearly defined and islets characterized using rigorous histopathology and immunolocalization examinations. Standardized procedures for processing human pancreas recovered from organ donors are described in part 1 of this series. The pancreas is processed into 3 main regions (head, body, tail) followed by transverse sections. Transverse sections from the pancreas head are further divided, as indicated based on size, and numbered alphabetically to denote subsections. This standardization allows for a complete cross sectional analysis of the head region including the uncinate region which contains islets composed primarily of pancreatic polypeptide cells to the tail region. The current report comprises part 2 of this series and describes the procedures used for serial sectioning and histopathological characterization of the pancreatic paraffin sections with an emphasis on islet endocrine cells, replication, and T-cell infiltrates. Pathology of pancreatic sections is intended to characterize both exocrine, ductular, and endocrine components. The exocrine compartment is evaluated for the presence of pancreatitis (active or chronic), atrophy, fibrosis, and fat, as well as the duct system, particularly in relationship to the presence of pancreatic intraductal neoplasia4. Islets are evaluated for morphology, size, and density, endocrine cells, inflammation, fibrosis, amyloid, and the presence of replicating or apoptotic cells using H&E and IHC stains. The final component described in part 2 is the provision of the stained slides as digitized whole slide images. The digitized slides are organized by case and pancreas region in an online pathology database creating a virtual biobank. Access to this online collection is currently provided to over 200 clinicians and scientists involved in type 1 diabetes research. The online database provides a means for rapid and complete data sharing and for investigators to select blocks for paraffin or frozen serial sections. PMID:22665223

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

    PubMed Central

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

    2011-01-01

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

  11. Centrifuge-operated specimen staining method and apparatus

    NASA Technical Reports Server (NTRS)

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

    1999-01-01

    A method of staining preselected, mounted specimens of either biological or nonbiological material enclosed within a staining chamber where the liquid staining reagents are applied and removed from the staining chamber using hypergravity as the propelling force. In the preferred embodiment, a spacecraft-operated centrifuge and method of diagnosing biological specimens while in orbit, characterized by hermetically sealing a shell assembly. The assembly contains slide stain apparatus with computer control therefor, the operative effect of which is to overcome microgravity, for example on board an International Space Station.

  12. Spectroscopic characterization of the binding mechanism of fluorescein and carboxyfluorescein in human serum albumin

    NASA Astrophysics Data System (ADS)

    Sulaiman, Saba A. J.; Kulathunga, H. Udani; Abou-Zied, Osama K.

    2015-03-01

    Fluorescein (FL) and some of its precursors have proven to be effective fluorescent tracers in pharmaceutical and medical applications owing to their high quantum yield of fluorescence in physiological conditions and their high membrane permeability. In order to protect FL from metabolic effects during the process of its delivery, human serum albumin (HSA) has been used as a carrier because of its compatibility with the human body. In the present work, we used spectroscopic methods to characterize the binding mechanisms of FL and one of its derivatives, 5(6)- carboxyfluorescein (CFL), in the HSA protein. The absorbance change of the two ligands (FL and CFL) was quantified as a function of the HSA concentration and the results indicate a moderate binding strength for the two ligands inside HSA (1.00 +/- 0.12 x 104 M-1). The quenching effect of FL(CFL) on the fluorescence intensity of W214 (the sole tryptophan in HSA) indicates that FL and CFL occupy Site I in the protein which is known to bind several hydrophobic drugs. By performing site-competitive experiments, the location of the ligands is determined to be similar to that of the anticoagulant drug warfarin. At higher ratios of [ligand]/[HSA], we observed an upward curvature in the Stern-Volmer plots which indicates that the ligands occupy more pockets in Site I, close to W214. Our results indicate that both ligands bind in HSA with a moderate strength that should not affect their release when used as fluorescent reporters. The chemical and physical identities of the two ligands are also preserved inside the HSA binding sites.

  13. Quantitative Analysis of Segmented Fluorescein Angiography Images for the Follow-up of Choroidal Neovascular Membrane

    PubMed Central

    Ghosh, Sambuddha; Haldar, Pampa; Ravindran, Prashanth; Chatterjee, Jyotirmoy; Paranjape, Sandeep V.; Bhaduri, Gautam

    2015-01-01

    Purpose: The aim of this study was to evaluate choroidal neovascular (CNV) lesions with fluorescein angiography (FA) and to identify quantitative parameters and correlate these parameters to treatment outcomes. Subjects and Methods: This institution based cross-sectional study evaluated 30 eyes with active sub-foveal predominantly classic CNV treated with bevacizumab. Pre- and post-injection segmented FA images were analyzed. Lesion area and CNV lesion were manually delineated. Outcome measure was the change 1-month after each injection in different intensity values (0–255 divided in eight regions A [lowest intensity] to H [highest intensity] on a linear scale) in lesion area, perimeter, greatest linear dimension (GLD), area, visual acuity (VA) and central macular thickness (CMT). Results: At month 3, statistically significant changes from baseline occurred in VA, CMT, lesion area, GLD and perimeter (P < 0.05 all comparisons). Change in CMT from baseline to 3 months postinjection was correlated with change in VA (P = 0.009, r = 0.469) and intensity regions B (P = 0.001, r = ?0.565), D (P = 0.001, r = 0.560), E (P = 0.035, r = 0.386). At month 3, change in intensity values 0–63 (A + B) was negatively correlated with CMT (P = 0.001, r = ?0.575) and lesion area (P = 0.019, r = ?0.427); change in intensity values 64–223 (C-G) was positively correlated with CMT (P = 0.000, r = 0.636) and lesion area (P = 0.002, r = 0.551). Conclusions: Decrease in area, GLD, perimeter and area with intensity ? 64 on segmented FA were associated with a favorable outcome of treatment. These parameters may be useful adjuncts to existing evaluation techniques during follow-up of CNV. PMID:25949076

  14. THE SUPRAVITAL STAINING OF VACCINE BODIES

    PubMed Central

    Cowdry, Edmund V.

    1922-01-01

    Vaccine bodies in living corneal cells may be specifically stained by the addition of a small quantity of brilliant cresyl blue 2 B to the physiological salt solution in which they are being observed. Their appearance by this method (Figs. 3 to 17) corresponds with that seen in fixed preparations (Figs. 22 to 42). Both lines of study reveal the existence of traces of similar material in unvaccinated corneal cells. As this increases in amount during the reaction, it behaves like an integral, cytoplasmic constituent of fluid consistency and shows no evidence of being endowed with any measure of independent vitality. The low grade of structural differentiation which it does exhibit, in living cells as well as in fixed tissues, is not suggestive of the presence within it of independent microorganisms. The material differs radically in its morphology and microchemical reactions from the granules observed by MacCallum and Oppenheimer in vaccine lymph. PMID:19868701

  15. Ultrafast tissue staining with chemical tags.

    PubMed

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

    2014-09-01

    Genetically encoded fluorescent proteins and immunostaining are widely used to detect cellular and subcellular structures in fixed biological samples. However, for thick or whole-mount tissue, each approach suffers from limitations, including limited spectral flexibility and lower signal or slow speed, poor penetration, and high background labeling, respectively. We have overcome these limitations by using transgenically expressed chemical tags for rapid, even, high-signal and low-background labeling of thick biological tissues. We first construct a platform of widely applicable transgenic Drosophila reporter lines, demonstrating that chemical labeling can accelerate staining of whole-mount fly brains by a factor of 100. Using viral vectors to deliver chemical tags into the mouse brain, we then demonstrate that this labeling strategy works well in mice. Thus this tag-based approach drastically improves the speed and specificity of labeling genetically marked cells in intact and/or thick biological samples. PMID:25157152

  16. Direct Blue 71 staining of proteins bound to blotting membranes.

    PubMed

    Hong, H Y; Yoo, G S; Choi, J K

    2000-03-01

    A sensitive staining method for protein blots using Direct Blue 71 is described. It is based on the selective binding of dye molecules to proteins in acidic solution and produces bluish violet colored bands. It is a simple and rapid procedure, involving only staining and rinsing steps that occur within 7 min. The sensitivity of this method is 5-10 ng of protein on nitrocellulose (NC) and 10-20 ng on polyvinylidene difluoride (PVDF), which is tenfold better than that of the commonly used Ponceau S staining. Moreover, the staining is reversible for subsequent immunostaining, without impairing immunoreactivity. To remove the dye from the developed bands, changes in pH and hydrophobicity of the solvent are required. Due to its sensitivity, rapidity, simplicity, and low cost, this stain may be more practical than other dye-based stains or metal-based stains for routine laboratory purposes. PMID:10768767

  17. Multi-class stain separation using independent component analysis

    NASA Astrophysics Data System (ADS)

    Trahearn, Nicholas; Snead, David; Cree, Ian; Rajpoot, Nasir

    2015-03-01

    Stain separation is the process whereby a full colour histology section image is transformed into a series of single channel images, each corresponding to a given stain's expression. Many algorithms in the field of digital pathology are concerned with the expression of a single stain, thus stain separation is a key preprocessing step in these situations. We present a new versatile method of stain separation. The method uses Independent Component Analysis (ICA) to determine a set of statistically independent vectors, corresponding to the individual stain expressions. In comparison to other popular approaches, such as PCA and NNMF, we found that ICA gives a superior projection of the data with respect to each stain. In addition, we introduce a correction step to improve the initial results provided by the ICA coefficients. Many existing approaches only consider separation of two stains, with primary emphasis on Haematoxylin and Eosin. We show that our method is capable of making a good separation when there are more than two stains present. We also demonstrate our method's ability to achieve good separation on a variety of different stain types.

  18. Further studies on the surface saccharides in Trichomonas vaginalis strains by fluorescein-conjugated lectins.

    PubMed

    Choroma?ski, L; Beat, D A; Nordin, J H; Pan, A A; Honigberg, B M

    1985-01-01

    Fluorescence emitted by individual cells of several Trichomonas vaginalis strains, nearly all of which were cloned, incubated with fluorescein-conjugated lectins in the absence (experimental) or presence (control) of inhibitory sugars, or else in phosphate-buffered saline alone (autofluorescence) was measured with a Leitz MPV Compact microfluorometer. Irrespective of whether the organisms were postfixed in formalin or glutaraldehyde, the relative fluorescence emitted by the cells was closely comparable, provided that appropriate neutral density filters were employed. However, autofluorescence was much higher for glutaraldehyde-fixed trichomonads. Therefore, although better preserved and more amenable to subsequent manipulations, such organisms were found unsuitable for use in "qualitative" titration of the fluorescence emitted by various strains. Provided that the necessary precautions were taken, comparable fluorescence readings were obtained with trichomonads affixed to glass slides by heat (41 degrees C, on a section spreader) or by a cytologic centrifuge (Cytospin 2). Large numbers of concanavalin A (Con A)-binding sites were present on organisms of all strains, irrespective of their virulence for human patients and as estimated by the subcutaneous mouse assay; these sites were shown with the aid of D-mannose to be mannose or mannose-related residues. More binding sites for soybean agglutinin (SBA) were found on the virulent than on avirulent strains. On the basis of inhibition experiments, the sugar residues mainly responsible for these differences appeared to be D-lactose residues. Similar differences were observed with Ricinus communis agglutinin Type I (RCA I), for which D-galactose was employed as the competing sugar. However, with two cloned strains the situation with regard to RCA I binding was reversed - more of the lectin bound to a mild than to a virulent strain. The results obtained with Ricinus communis Type II agglutinin (RCA II) were often similar to those noted for RCA I; however, in most instances the inhibition with N-acetyl-D-galactosamine (GalNAc) was lower. Furthermore, the results noted with the GalNAc-specific agglutinins from Dolichus biflorus and Helix pomatia suggested that only very few GalNAc residues were available for binding on the surfaces of all T. vaginalis strains examined in the course of this study. Statistical analyses of fluorescence emitted by four clones of each, Balt 42 (virulent) and JH31A (avirulent) T. vaginalis strain upon incubation with Con A and SBA revealed homogeneity of these strains with regard to the number of the specific surface saccharide residues, D-mannose and D-lactose.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:3927600

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

    PubMed Central

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

    2011-01-01

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

  20. Visible luminescence from silicon wafers subjected to stain etches

    NASA Technical Reports Server (NTRS)

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

    1992-01-01

    Etching of Si in a variety of solutions is known to cause staining. These stain layers consist of porous material similar to that produced by anodic etching of Si in HF solutions. In this work, photoluminescence peaked in the red from stain-etched Si wafers of different dopant types, concentrations, and orientations produced in solutions of HF:HNO3:H2O was observed. Luminescence is also observed in stain films produced in solutions of NaNO2 in HF, but not in stain films produced in solutions of CrO3 in HF. The luminescence spectra are similar to those reported recently for porous Si films produced by anodic etching in HF solutions. However, stain films are much easier to produce, requiring no special equipment.

  1. Modified Genta triple stain for identifying Helicobacter pylori.

    PubMed Central

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

    1999-01-01

    AIM: To evaluate whether lead nitrate could replace uranyl nitrate in the Genta stain for H pylori without sacrificing the advantages of the triple stain (Steiner silver impregnation combined with Alcian blue and haematoxylin/eosin (H&E)). METHODS: A comparison was made in 16 specimens between the original triple stain and the revised version. One pathologist evaluated all sections. RESULTS: Direct substitution of lead nitrate for uranium nitrate produced well stained organisms without interfering with H&E or Alcian blue staining. No difference was found in the ability to identify bacteria in 11 cases with H pylori density of 1 or 2 (on a scale of 0 to 5). CONCLUSIONS: The potential chemical and radiological hazards associated with uranium nitrate can be eliminated by using lead nitrate without sacrificing the advantages obtained by using the triple stain. Images PMID:10655993

  2. Vascular invasion of colorectal carcinoma readily visible with certain stains

    Microsoft Academic Search

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

    1992-01-01

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

  3. Effects of ?-galactosidase digestion on lectin staining in human pancreas

    Microsoft Academic Search

    N. Ito; K. Nishi; M. Nakajima; Y. Okamura; T. Hirota

    1988-01-01

    Effects of a-galactosidase (from green coffee beans) digestion on lectin staining were examined in formalin-fixed, paraffin-embedded human pancreatic tissues from individuals of blood-group B and AB. Digestion with the enzyme resulted in almost complete loss of Griffonia simplicifolia agglutinin I-B4(GSAI-B4) staining in the acinar cells with concomitant appearance of Ulex europaeus agglutinin-I(UEA-I) staining in the corresponding cells. In addition, reactivity

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

    PubMed

    Biesemeier, Antje; Schraermeyer, Ulrich; Eibl, Oliver

    2011-07-01

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

  5. Fluorescein analogues inhibit SecA ATPase: the first sub-micromolar inhibitor of bacterial protein translocation.

    PubMed

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

    2012-04-01

    SecA is a central component of the general secretion system that is essential for bacterial growth and thus an ideal target for antimicrobial agents. A series of fluorescein analogues were first screened against the ATPase activity using the truncated unregulated SecA catalytic domain. Rose bengal (RB) and erythrosin B (EB) were found to be potent inhibitors SecA with IC(50) values of 0.5 ?M and 2 ?M, respectively. RB and EB inhibit the catalytic SecA ATPase more effectively than the F(1) F(0) -proton ATPase. We used three assays to test the effect of these compounds on full-length SecA ATPase: in solution (intrinsic ATPase), in membrane preparation, and translocation ATPase. RB and EB show the following trend in terms of IC(50) values: translocation ATPasefluorescein analogues in inhibiting the truncated SecA ATPase correlates with their ability to inhibit the biologically relevant protein translocation activity of SecA. The in vitro translocation of proOmpA precursors into membrane vesicles is strongly inhibited by RB with IC(50) values of approximately 0.25 ?M, making RB the most potent inhibitor of SecA ATPase and SecA-dependent protein translocation reported thus far. The ability of these compounds to inhibit SecA also directly translates into antibacterial effects. Our findings show the value of fluorescein analogues as probes for mechanistic studies of SecA functions and for the potential development of new antimicrobial agents with SecA as the target. PMID:22354575

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

    PubMed Central

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

    2014-01-01

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

  7. Sihler's whole mount nerve staining technique: a review

    PubMed Central

    Mu, L; Sanders, I

    2009-01-01

    Sihler's stain is a whole mount nerve staining technique that renders other soft tissue translucent or transparent while staining the nerves. It permits mapping of entire nerve supply patterns of organs, skeletal muscles, mucosa, skin, and other structures after the specimens are fixed in neutralized formalin, macerated in potassium hydroxide, decalcified in acetic acid, stained in Ehrlich's hematoxylin, destained in acetic acid, and cleared in glycerin. The unique advantage of Sihler's stain over other anatomical methods is that all the nerves within the stained specimen can be visualized in their three-dimensional positions. To date, Sihler's stain is the best tool for demonstrating the precise intramuscular branching and distribution patterns of skeletal muscles, which are important not only for anatomists, but also for physiologists and clinicians. Advanced knowledge of the neural structures within mammalian skeletal muscles is critical for understanding muscle functions, performing electrophysiological experiments and developing novel neurosurgical techniques. In this review, Sihler's stain is described in detail and its use in nerve mapping is surveyed. Special emphasis is placed on staining procedures and troubleshooting, strengths and limitations, applications, major contributions to neuroscience, physiological and clinical significance, and areas for further technical improvement that deserve future research. PMID:19572223

  8. Fluorescein analogue xanthene-9-carboxylic acid: a transition-metal-free CO releasing molecule activated by green light.

    PubMed

    Antony, Lovely Angel Panamparambil; Slanina, Tomáš; Šebej, Peter; Šolomek, Tomáš; Klán, Petr

    2013-09-01

    6-Hydroxy-3-oxo-3H-xanthene-9-carboxylic acid is introduced as the first transition-metal-free carbon monoxide releasing molecule activated by visible light (photoCORM). This water-soluble fluorescein analogue releases carbon monoxide in both water and methanol upon irradiation at 500 nm. When selectively irradiated in the presence of hemoglobin (Hb) under physiological conditions, released CO is quantitatively trapped to form carboxyhemoglobin (COHb). The reaction progress can be accurately monitored by characteristic absorption and emission properties of the reactants and products. PMID:23957602

  9. SYPRO Orange and SYPRO Red Protein Gel Stains: One-Step Fluorescent Staining of Denaturing Gels for Detection of Nanogram Levels of Protein

    Microsoft Academic Search

    Thomas H. Steinberg; Laurie J. Jones; Richard P. Haugland; Victoria L. Singer

    1996-01-01

    We have developed two new fluorescent dyes, SYPRO Orange protein gel stain and SYPRO Red protein gel stain, to detect proteins in electrophoretic gels. Stained protein bands can be excited by ultraviolet light at ?300 nm, or at visible wavelengths, with excitation maxima of 472 nm for the Orange stain and 547 nm for the Red stain. Detection can be

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

    NASA Astrophysics Data System (ADS)

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

    2001-12-01

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

  11. Control of Listeria monocytogenes on cooked cured ham by formulation with a lactate-diacetate blend and surface treatment with lauric arginate.

    PubMed

    Stopforth, J D; Visser, D; Zumbrink, R; van Dijk, L; Bontenbal, E W

    2010-03-01

    Ready-to-eat (RTE) meat products have been identified as a significant source of listeriosis in humans in the United States. Meat processors in the United States are required to use one of three alternatives to control L. monocytogenes in RTE meats: (i) a postlethality inactivation treatment along with a L. monocytogenes growth inhibitor; (ii) a postlethality inactivation treatment or a growth inhibitor; or (iii) sanitation measures and intensive testing. Lauric arginate (LAE) has been proposed as an effective postlethality inactivation treatment. The present study was conducted to investigate the antimicrobial effect of a lactate-diacetate blend in the formulation combined with surface application of LAE on cooked cured ham inoculated with L. monocytogenes, vacuum packaged, and stored at 4 degrees C for up to 90 days. The treatments evaluated were (i) control ham with no added antimicrobials (control); (ii) ham formulated with 1.68% potassium lactate and 0.12% sodium diacetate (PLSD); (iii) control ham with 0.07% LAE as a surface treatment (LAE); and (iv) ham formulated with PLSD and LAE surface treatment (sprayed in bag and distributed across meat surface during vacuum packing) (PLSD + LAE). Use of only LAE as a surface treatment resulted in an initial 1-log CFU/g reduction in levels of L. monocytogenes on ham; however, this reduction only delayed the growth of the pathogen to 8 log CFU/g by 12 days when compared with the control ham without added antimicrobials. Use of PLSD in the formulation of ham resulted in a complete inhibition of L. monocytogenes throughout storage. The combination of PLSD in the formulation and a surface treatment with LAE resulted in an initial 0.7-log CFU/g reduction of the pathogen on ham and complete inhibition of the pathogen at the reduced level throughout storage. Formulation of ham with a lactate-diacetate blend combined with lauric arginate as a surface treatment will allow RTE meat processors to effectively achieve alternative 1 status, as designated by the U.S. Department of Agriculture Food Safety and Inspection Service, in their facilities. PMID:20202344

  12. Klutts 4/2004 Silver Staining of Protein Gels

    E-print Network

    Doering, Tamara

    Klutts 4/2004 1 Silver Staining of Protein Gels Overview: Silver staining is much more sensitive than Coomassie, typically you can see 10-50 ng of a protein. It does vary, however, with the glycosylation and physical properties of the protein. (Protocol from Jeff Brodsky to Tamara.) Materials (enough

  13. Development of an affordable dye-stained microalbuminuria screening test

    Microsoft Academic Search

    Pierrot Lundimu Tugirimana; Joris R. Delanghe

    2008-01-01

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

  14. Unusual indelible enamel staining following fixed appliance treatment.

    PubMed

    Hodges, S J; Spencer, R J; Watkins, S J

    2000-12-01

    Two cases are described of indelible enamel staining following fixed appliance therapy. The acquired pigmentation occurred in patients with an identifiable enamel defect prior to treatment. The interaction of factors to cause the staining is discussed and it's prevention in future cases highlighted. Subsequent restoration of the affected teeth is shown. PMID:11099567

  15. Alcian Blue Alizarin Red Skeletal Staining October 2003

    E-print Network

    De Robertis, Eddy M.

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

  16. The effect of selected staining techniques on bull sperm morphometry.

    PubMed

    Banaszewska, Dorota; Andraszek, Katarzyna; Czubaszek, Magdalena; Biesiada-Drzazga, Barbara

    2015-08-01

    Sperm morphometry has some value as an indicator of reproductive capacity in males. In laboratory practice a variety of slide-staining methods are used during morphological evaluation of semen to predict male fertility. The aim of this study was to determine the effect of staining of semen using four different techniques on the morphometry of the bull sperm cell. The material for the study consisted of semen collected from test bulls of the Black-and-White variety of Holstein-Friesians. The results obtained in the study indicate differences in the dimensions of bull sperm heads when different slide staining techniques were used. The most similar results for sperm head dimensions were obtained in the case of SpermBlue(®) and eosin+gentian violet complex, although statistically significant differences were found between all the staining techniques. Extreme values were noted for the other staining techniques - lowest for the Papanicolaou and highest for silver nitrate, which may indicate more interference in the cell by the reagents used in the staining process. However, silver nitrate staining was best at identifying the structures of the sperm cell. Hence it is difficult to determine which of the staining methods most faithfully reveals the dimensions and shape of the bull sperm. PMID:26149220

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

    Microsoft Academic Search

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

    1996-01-01

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

  18. Optimization of size, morphology and colloidal stability of fluorescein dye-doped silica NPs for application in immunoassays.

    PubMed

    Nooney, Robert I; McCormack, Eoin; McDonagh, Colette

    2012-12-01

    Fluorescent nanoparticle (NP) labels are of great interest for point-of-care medical diagnostics where high fluorescence signals combined with low limits of detection are required. In this work, hydrophilic and hydrophobic fluorescein dye derivatives were covalently doped into silica NPs. The NPs were prepared in a range of sizes from 16 to 80 nm using both ternary and quaternary microemulsion methods where the diameter varied linearly with changes in the water to surfactant ratio. The morphology and colloidal stability of the NPs were characterised using transmission electron microscopy and photon correlation spectroscopy; NPs doped with hydrophobic fluorescein dye were significantly smaller and more polydispersed. Optical properties including absorption, fluorescence and quantum efficiency were also determined. Representative NPs from each microemulsion method (ternary, Ø = 25 nm and quaternary, Ø = 80 nm) were tested as labels in a fluorescence based immunoassay for the detection of human IgG and human chorionic gonadotropin. Both sets of nanoparticle assays showed lower limits of detection and better coefficients of variance than a free dye label with good day to day reproducibility. The optimal surface coverage of detection antibody was also found to depend on the size of the nanoparticle. PMID:22868474

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

    NASA Astrophysics Data System (ADS)

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

    2010-12-01

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

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

    PubMed Central

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

    1992-01-01

    Three model biomembrane systems, monolayers, micelles, and vesicles, have been used to study the influence of chemical and physical variables of hapten presentation at membrane interfaces on antibody binding. Hapten recognition and binding were monitored for the anti-fluorescein monoclonal antibody 4-4-20 generated against the hapten, fluorescein, in these membrane models as a function of fluorescein-conjugated lipid architecture. Specific recognition and binding in this system are conveniently monitored by quenching of fluorescein emission upon penetration of fluorescein into the antibody's active site. Lipid structure was shown to play a large role in affecting antibody quenching. Interestingly, the observed degrees of quenching were nearly independent of the lipid membrane model studied, but directly correlated with the chemical structure of the lipids. In all cases, the antibody recognized and quenched most efficiently a lipid based on dioctadecylamine where fluorescein is attached to the headgroup via a long, flexible hydrophilic spacer. Dipalmitoyl phosphatidylethanolamine containing a fluorescein headgroup demonstrated only partial binding/quenching. Egg phosphatidylethanolamine with a fluorescein headgroup showed no susceptibility to antibody recognition, binding, or quenching. Formation of two-dimensional protein domains upon antibody binding to the fluorescein-lipids in monolayers is also presented. Chemical and physical requirements for these antibody-hapten complexes at membrane surfaces have been discussed in terms of molecular dynamics simulations based on recent crystallographic models for this antibody-hapten complex (Herron et al., 1989. Proteins Struct. Funct. Genet. 5:271-280). Images FIGURE 7 FIGURE 8 PMID:1420916

  1. Western Blot of Stained Proteins from Dried Polyacrylamide Gels

    NASA Technical Reports Server (NTRS)

    Gruber, Claudia; Stan-Lotter, Helga

    1996-01-01

    Western blotting of proteins is customarily performed following their separation on polyacrylamide gels, either prior to staining (1) or, as recently reported, following staining (2). We describe here Western blotting with stained gels, which had been dried and some of which had been stored for years. This procedure permits immunological analysis of proteins, to which antisera may have become available only later, or where the application of newly developed sensitive detection methods is desired. Once rehydration of the gels is achieved, proteins can be-transferred to blotting membranes by any appropriate protocol. Proteins stained with Coomassie Blue have to be detected with a non-chromogenic method, such as the film-based enhanced chemiluminescence (ECL)2) procedure (3). Silver stained proteins, which transfer in the colorless form, may be visualized by any detection method, although, because of the usually very low amounts of proteins, detection by ECL is preferable. Blotting of stained proteins from rehydrated gels is as rapid and as quantitative as from freshly prepared gels, in contrast to blotting from wet stained gels, which requires extensive washing and results in low transfer efficiency (2). Together with a photographic record of the gel pattern, unambiguous identification of immunoreactive proteins from complex mixtures is possible. Some further applications of this work are discussed.

  2. Self-assembled monolayers of 6-mercapto-1-hexanol and mercapto- n-hexyl-poly(dT)18-fluorescein on polycrystalline gold surfaces: An electrochemical impedance spectroscopy study

    Microsoft Academic Search

    Joel Rivera-Gandía; Carlos R. Cabrera

    2007-01-01

    The following study presents the analysis of electrochemically induced conformational changes of self-assembled single-stranded DNA (mercapto-n-hexyl-poly(dT)18-fluorescein, 18TFAM) on polished polycrystalline Au electrodes. This was compared to a self-assembled monolayer of 6-mercapto-1-hexanol (MCH). Double layer capacitance (Cdl) was determined by electrochemical impedance spectroscopy (EIS) with electrochemical perturbation from 0.0V to 0.7V (vs. Ag\\/AgCl) for bare Au, Au\\/mercaptohexanol (Au\\/MCH), and Au\\/S-(CH2)6-poly(dT)18-fluorescein (Au\\/18TFAM)

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

    NASA Astrophysics Data System (ADS)

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

    2012-03-01

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

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

    PubMed

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

    2010-04-16

    PEGylated PAMAM conjugated fluorescein-doped magnetic silica nanoparticles (PEGylated PFMSNs) have been synthesized for evaluating their ability across the blood-brain barrier (BBB) and distribution in rat brain. The obtained nanoparticles were characterized by transmission electron microscopy (TEM), thermal gravimetry analyses (TGA), zeta potential (zeta-potential) titration, and X-ray photoelectron spectroscopy (XPS). The BBB penetration and distribution of PEGylated PFMSNs and FMSNs in rat brain were investigated not only at the cellular level with Confocal laser scanning microscopy (CLSM), but also at the subcellular level with transmission electron microscopy (TEM). The results provide direct evidence that PEGylated PFMSNs could penetrate the BBB and spread into the brain parenchyma. PMID:20206605

  5. Accurate measurement of avidin and streptavidin in crude biofluids with a new, optimized biotin-fluorescein conjugate.

    PubMed

    Kada, G; Falk, H; Gruber, H J

    1999-03-14

    A new biotin-fluorescein conjugate with an ethylene diamine spacer was found to be the first fluorescent biotin derivative which truly mimicked d-biotin in terms of high affinity, fast association, and non-cooperative binding to avidin and streptavidin tetramers. These exceptional properties were attributed to the small size/length of the new ligand since all larger/longer biotin derivatives are known for their mutual steric hindrance and anti-cooperative binding in 4:1 complexes with avidin and streptavidin tetramers. Specific binding of the new biotin-fluorescein conjugate towards avidin and streptavidin was accompanied by 84-88% quenching of ligand fluorescence. In the accompanying study this effect was used for rapid estimation of avidin and streptavidin in a new 'single tube assay'. In the present study the strong quenching effect was utilized to accurately monitor stoichiometric titration of biotin-binding sites in samples with >/=200 pM avidin or streptavidin. The concentration was calculated from the consumption of fluorescent ligand up to the distinct breakpoint in the fluorescence titration profile which was marked by the abrupt appearance of strongly fluorescent ligands which were in excess. Due to this protocol the assay was not perturbed by background fluorescence or coloration in the unknown samples. The new fluorescence titration assay is particularly suited for quick checks on short notice because getting started only means to thaw an aliquot of a standardized stock solution of fluorescent ligand. No calibration is required for the individual assay and the ligand stock solution needs to be restandardized once per week (or once per year) when stored at -25 degrees C (or at -70 degrees C, respectively). PMID:10082985

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

    PubMed Central

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

    2001-01-01

    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.?? PMID:11316723

  7. Immunohistochemical staining for ranaviruses Introduction: Ranaviruses negatively impact amphibian populations

    E-print Network

    Gray, Matthew

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

  8. Klutts 4/2004 Coomassie Staining of Protein Gels

    E-print Network

    Doering, Tamara

    Klutts 4/2004 1 Coomassie Staining of Protein Gels Overview: The laboratory typically uses the Bio if you are staining a gel with a lot of protein, as it is not quite as sensitive on re-use. 3. You canRad catalog number 161-0786. Bio-Safe Coomassie: 1. Wash your gel for 10 minutes in water to remove SDS

  9. Immunohistochemical CD3 staining detects additional patients with celiac disease

    PubMed Central

    Mubarak, Amani; Wolters, Victorien M; Houwen, Roderick HJ; ten Kate, Fiebo JW

    2015-01-01

    AIM: To investigate whether performing immunohistochemical CD3 staining, in order to improve the detection of intra-epithelial lymphocytosis, has an additional value in the histological diagnosis of celiac disease. METHODS: Biopsies obtained from 159 children were stained by hematoxylin and eosin (HE) and evaluated using the Marsh classification. CD3 staining was subsequently evaluated separately and independently. RESULTS: Differences in evaluation between the routine HE sections and CD3 staining were present in 20 (12.6%) cases. In 10 (6.3%) patients the diagnosis of celiac disease (Marsh II and III) changed on examination of CD3 staining: in 9 cases, celiac disease had initially been missed on the HE sections, while 1 patient had been over-diagnosed on the routine sections. In all patients, the final diagnosis based on CD3 staining, was concordant with serological results, which was not found previously. In the other 10 (12.3%) patients, the detection of sole intra-epithelial lymphocytosis (Marsh?I) improved. Nine patients were found to have Marsh?I?on CD3 sections, which had been missed on routine sections. Interestingly, the only patient with negative serology had Giardiasis. Finally, in 1 patient with negative serology, in whom Marsh?I?was suspected on HE sections, this diagnosis was withdrawn after evaluation of the CD3 sections. CONCLUSION: Staining for CD3 has an additional value in the histological detection of celiac disease lesions, and CD3 staining should be performed when there is a discrepancy between serology and the diagnosis made on HE sections.

  10. 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

  11. Influence of Local Treatments with Capsaicin or Allyl Isothiocyanate in the Sensitization Phase of a Fluorescein-Isothiocyanate-Induced Contact Sensitivity Model

    Microsoft Academic Search

    Takashi Maruyama; Hiromi Iizuka; Yuki Tobisawa; Takahiro Shiba; Tomoko Matsuda; Kohta Kurohane; Yasuyuki Imai

    2007-01-01

    Background: In fluorescein isothiocyanate (FITC)-induced contact hypersensitivity models, dibutyl phthalate has been empirically used as a solvent ingredient. We have demonstrated that dibutyl phthalate has an adjuvant effect through the facilitation of trafficking FITC-presenting dendritic cells (DC) from the skin to draining lymph nodes. Here we investigated the effects of local pretreatment with substances that are capable of desensitizing sensory

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

    Microsoft Academic Search

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

    1993-01-01

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

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

    Microsoft Academic Search

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

    2003-01-01

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

  14. Immunohistochemical staining of radixin and moesin in prostatic adenocarcinoma

    PubMed Central

    2011-01-01

    Background Some members of the Protein 4.1 superfamily are believed to be involved in cell proliferation and growth, or in the regulation of these processes. While the expression levels of two members of this family, radixin and moesin, have been studied in many tumor types, to our knowledge they have not been investigated in prostate cancer. Methods Tissue microarrays were immunohistochemically stained for either radixin or moesin, with the staining intensities subsequently quantified and statistically analyzed using One-Way ANOVA or nonparametric equivalent with subsequent Student-Newman-Keuls tests for multiple comparisons. There were 11 cases of normal donor prostates (NDP), 14 cases of benign prostatic hyperplasia (BPH), 23 cases of high-grade prostatic intraepithelial neoplasia (HGPIN), 88 cases of prostatic adenocarcinoma (PCa), and 25 cases of normal tissue adjacent to adenocarcinoma (NAC) analyzed in the microarrays. Results NDP, BPH, and HGPIN had higher absolute staining scores for radixin than PCa and NAC, but with a significant difference observed between only HGPIN and PCa (p = < 0.001) and HGPIN and NAC (p = 0.001). In the moesin-stained specimens, PCa, NAC, HGPIN, and BPH all received absolute higher staining scores than NDP, but the differences were not significant. Stage 4 moesin-stained PCa had a significantly reduced staining intensity compared to Stage 2 (p = 0.003). Conclusions To our knowledge, these studies represent the first reports on the expression profiles of radixin and moesin in prostatic adenocarcinoma. The current study has shown that there were statistically significant differences observed between HGPIN and PCa and HGPIN and NAC in terms of radixin expression. The differences in the moesin profiles by tissue type were not statistically significant. Additional larger studies with these markers may further elucidate their potential roles in prostatic neoplasia progression. PMID:21235778

  15. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...Painted, colored or stained glass windows for religious institutions. 10...Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are...

  16. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...Painted, colored or stained glass windows for religious institutions. 10...Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are...

  17. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...Painted, colored or stained glass windows for religious institutions. 10...Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are...

  18. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...Painted, colored or stained glass windows for religious institutions. 10...Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are...

  19. 19 CFR 10.52 - Painted, colored or stained glass windows for religious institutions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...Painted, colored or stained glass windows for religious institutions. 10...Painted, colored or stained glass windows for religious institutions. When painted, colored, or stained glass windows or parts thereof, are...

  20. Methods of biological dosimetry employing chromosome-specific staining

    DOEpatents

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

    2000-01-01

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Further, methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. Still further, the invention provides for automated means to detect and analyse chromosomal abnormalities.

  1. Methods And Compositions For Chromosome-Specific Staining

    DOEpatents

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

    2003-08-19

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods are provided to disable the hybridization capacity of shared, high copy repetitive sequences and/or remove such sequences to provide for useful contrast. Still further methods are provided to produce chromosome-specific staining reagents which are made specific to the targeted chromosomal material, which can be one or more whole chromosomes, one or more regions on one or more chromosomes, subsets of chromosomes and/or the entire genome. Probes and test kits are provided for use in tumor cytogenetics, in the detection of disease related loci, in analysis of structural abnormalities, such as translocations, and for biological dosimetry. Further, methods and prenatal test kits are provided to stain targeted chromosomal material of fetal cells, including fetal cells obtained from maternal blood. Still further, the invention provides for automated means to detect and analyse chromosomal abnormalities.

  2. Chromosome-specific staining to detect genetic rearrangements

    DOEpatents

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

    2013-04-09

    Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyzes. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML) and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar but genetically different diseases, and for many prognostic and diagnostic applications.

  3. A staining protocol for identifying secondary compounds in Myrtaceae1

    PubMed Central

    Retamales, Hernan A.; Scharaschkin, Tanya

    2014-01-01

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

  4. Specific cytochemical stains in the image analysis of subcellular organelles.

    PubMed

    Cornelis, A; Van Meerbeek, D; Nyssen, M; Roels, F

    1985-12-01

    This paper describes our work concerning densitometry and morphometry of subcellular structures in thin sections. The techniques of automatic image analysis were applied to light and electron microscopic observations of enzymatically stained lysosomes, renal brush borders and mitochondria (in human and rat kidney) and peroxisomes (in human liver). To obtain significant measurements of the enzymatic activity, specific staining techniques were developed and applied, including an improved staining of acid phosphatase for lysosomes. Optical densities were obtained by videodensitometry and electron densities of peroxisomes were obtained by digitizing and processing scanning transmission electron microscopic images. In subsequent steps, delineations and parameter estimation are performed by software. Included was an examination of delineation techniques, which showed improved results from the use of a newly developed local boundary search algorithm. The combination of these techniques was used to study changes in peroxisome and lysosome compartment in liver and kidney, some results of which are also reported. PMID:2418853

  5. News from the Biological Stain Commission, no. 16.

    PubMed

    Lyon, H O

    2015-04-01

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

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

    PubMed Central

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

    2009-01-01

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

  7. Competition-based cellular peptide binding assays for 13 prevalent HLA class I alleles using fluorescein-labeled synthetic peptides.

    PubMed

    Kessler, Jan H; Mommaas, Bregje; Mutis, Tuna; Huijbers, Ivo; Vissers, Debby; Benckhuijsen, Willemien E; Schreuder, Geziena M Th; Offringa, Rienk; Goulmy, Els; Melief, Cornelis J M; van der Burg, Sjoerd H; Drijfhout, Jan W

    2003-02-01

    We report the development, validation, and application of competition-based peptide binding assays for 13 prevalent human leukocyte antigen (HLA) class I alleles. The assays are based on peptide binding to HLA molecules on living cells carrying the particular allele. Competition for binding between the test peptide of interest and a fluorescein-labeled HLA class I binding peptide is used as read out. The use of cell membrane-bound HLA class I molecules circumvents the need for laborious biochemical purification of these molecules in soluble form. Previously, we have applied this principle for HLA-A2 and HLA-A3. We now describe the assays for HLA-A1, HLA-A11, HLA-A24, HLA-A68, HLA-B7, HLA-B8, HLA-B14, HLA-B35, HLA-B60, HLA-B61, and HLA-B62. Together with HLA-A2 and HLA-A3, these alleles cover more than 95% of the Caucasian population. Several allele-specific parameters were determined for each assay. Using these assays, we identified novel HLA class I high-affinity binding peptides from HIVpol, p53, PRAME, and minor histocompatibility antigen HA-1. Thus these convenient and accurate peptide-binding assays will be useful for the identification of putative cytotoxic T lymphocyte epitopes presented on a diverse array of HLA class I molecules. PMID:12559627

  8. Banding patterns in newt chromosomes by the giemsa stain

    Microsoft Academic Search

    Irma Nardi; Matilde Ragghianti; Giorgio Mancino

    1973-01-01

    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

  9. Nuclear stains with soluble metachrome metal mordant dye lakes

    Microsoft Academic Search

    R. D. Lillie; P. Pizzolato; P. T. Donaldson

    1976-01-01

    Following our study on the effect of deoxyribonucleic acid (DNA) extraction on nuclear staining with soluble metal mordant dye lakes covering 29 dye lakes we chose a series of lakes representing the three groups: (1) readily prevented by DNA removal, (2) weakened by DNA extraction but not prevented, (3) unaffected by DNA removal, for application of other endgroup blockade reactions.

  10. ENHANCED DAPI STAINING FOR CRYPTOSPORIDIUM IN WATER SAMPLES

    EPA Science Inventory

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

  11. 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

  12. A Genetic Algorithm for Grammars James Anderson and Joe Staines

    E-print Network

    Goldschmidt, Christina

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

  13. Research on Textile Stain Detection Technique Based on Machine Vision

    Microsoft Academic Search

    Su Chen; Wang Huang; Lin Qing; Zhao Da-xing; Zhu Jin-lei

    2009-01-01

    The automatic detection of medical textile stains has always been a hot spot. An automatic detection method is proposed based on machine vision, which can solve the technical difficult problem of automatic detection technique efficiently. The procedure of the textile image processing has been researched, and the emphasis should be put on the image preprocessing and edge extraction. The experiments

  14. Gram's Stain Does Not Cross the Bacterial Cytoplasmic Membrane.

    PubMed

    Wilhelm, Michael J; Sheffield, Joel B; Sharifian Gh, Mohammad; Wu, Yajing; Spahr, Christian; Gonella, Grazia; Xu, Bolei; Dai, Hai-Lung

    2015-07-17

    For well over a century, Hans Christian Gram's famous staining protocol has been the standard go-to diagnostic for characterizing unknown bacteria. Despite continuous and ubiquitous use, we now demonstrate that the current understanding of the molecular mechanism for this differential stain is largely incorrect. Using the fully complementary time-resolved methods: second-harmonic light-scattering and bright-field transmission microscopy, we present a real-time and membrane specific quantitative characterization of the bacterial uptake of crystal-violet (CV), the dye used in Gram's protocol. Our observations contradict the currently accepted mechanism which depicts that, for both Gram-negative and Gram-positive bacteria, CV readily traverses the peptidoglycan mesh (PM) and cytoplasmic membrane (CM) before equilibrating within the cytosol. We find that not only is CV unable to traverse the CM but, on the time-scale of the Gram-stain procedure, CV is kinetically trapped within the PM. Our results indicate that CV, rather than dyes which rapidly traverse the PM, is uniquely suited as the Gram stain. PMID:25879387

  15. PREFERENTIAL STAINING OF NUCLEIC ACIDCONTAINING STRUCTURES FOR ELECTRON MICROSCOPY

    Microsoft Academic Search

    H. E. Huxley; G. ZUBAY

    1961-01-01

    Oriented fibres of extracted nuclcohistone were employed as test material in a study of satisfactory fixation, embedding, and staining methods for structures containing a high proportion of nucleic acid. Fixation in buffered osmium tctroxide solution at pH 6, con- taining l0 -2 M Ca ++, and embedding in Araldite enabled sections of the fibres to be cut in which the

  16. ANEUPLOIDY TEST DEVELOPMENT: KINETOCHORE STAINING IN MAMMALIAN SYSTEMS

    EPA Science Inventory

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

  17. Biocytin staining of glia and neurons in brain slices.

    PubMed

    Kang, Jian

    2014-09-01

    This protocol describes the use of biocytin to visualize and distinguish the morphology of glia and neurons in rat brain slices. Patch pipettes are used to load biocytin into different cell types. The slices are subsequently fixed, stained, and mounted in preparation for imaging. PMID:25183822

  18. MP 33308 Multiplexed ProteomicsTM Transmembrane Protein Gel Stain Kit Product Information

    E-print Network

    Lebendiker, Mario

    MP 33308 Multiplexed ProteomicsTM Transmembrane Protein Gel Stain Kit Product Information Storage upon receipt: Protein Gel Stains (Components A and B) · Room temperature · Protect from light gel stain · 280, 450/610 nm for SYPRO Ruby protein gel stain Quick Facts Revised: 05­August­2003

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

    PubMed

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

    2014-11-14

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

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

    PubMed Central

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

    2010-01-01

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

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

    Microsoft Academic Search

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

    1989-01-01

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

  2. Drying and curing of stains and lacquers used in furniture finishing 1 DRYING AND CURING OF STAINS AND LACQUERS USED IN

    E-print Network

    Stokes, Yvonne

    Drying and curing of stains and lacquers used in furniture finishing 1 DRYING AND CURING OF STAINS AND LACQUERS USED IN FURNITURE FINISHING Y.M. Stokes1 and P. Pendleton2 1. Problem description Nexus Pty Ltd. The furniture is finished with a stain and two lacquer coatings which give it a semi-gloss surface. In 1997

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

    PubMed

    Janchaysang, Suwatwong; Sumriddetchkajorn, Sarun; Buranasiri, Prathan

    2012-10-10

    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

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

    PubMed

    Hochstrasser, D F; Merril, C R

    1988-01-01

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

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

    SciTech Connect

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

    2010-07-02

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

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

    NASA Astrophysics Data System (ADS)

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

    2013-08-01

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

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

    NASA Astrophysics Data System (ADS)

    Hara, Shigemitsu; Tanoue, Eiichiro

    1989-11-01

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

  8. Lectins stain cells differentially in the coral, Montipora capitata

    USGS Publications Warehouse

    Work, Thierry M.; Farah, Yael

    2014-01-01

    A limitation in our understanding of coral disease pathology and cellular pathogenesis is a lack of reagents to characterize coral cells. We evaluated the utility of plant lectins to stain tissues of a dominant coral, Montipora capitata, from Hawaii. Of 22 lectins evaluated, nine of these stained structures in the upper or basal body wall of corals. Specific structures revealed by lectins that were not considered distinct or evident on routine hematoxylin and eosin sections of coral tissues included apical and basal granules in gastrodermis and epidermis, cnidoglandular tract and actinopharynx cell surface membranes, capsules of mature holotrichous isorhizas, and perivitelline and periseminal cells. Plant lectins could prove useful to further our understanding of coral physiology, anatomy, cell biology, and disease pathogenesis.

  9. Identification of active fluorescence stained bacteria by Raman spectroscopy

    NASA Astrophysics Data System (ADS)

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

    2008-04-01

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

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

    NSDL National Science Digital Library

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

  11. Genetic Variants Associated with Port-Wine Stains

    PubMed Central

    Wooderchak-Donahue, Whitney; Tan, Oon T.; Margraf, Rebecca; Stevenson, David A.; Grimmer, J. Fredrik; Bayrak-Toydemir, Pinar

    2015-01-01

    Background Port-wine stains (PWS) are capillary malformations, typically located in the dermis of the head and neck, affecting 0.3% of the population. Current theories suggest that port-wine stains are caused by somatic mutations that disrupt vascular development. Objectives Understanding PWS genetic determinants could provide insight into new treatments. Methods Our study used a custom next generation sequencing (NGS) panel and digital polymerase chain reaction to investigate genetic variants in 12 individuals with isolated port-wine stains. Importantly, affected and healthy skin tissue from the same individual were compared. A subtractive correction method was developed to eliminate background noise from NGS data. This allowed the detection of a very low level of mosaicism. Results A novel somatic variant GNAQ, c.547C>G, p.Arg183Gly was found in one case with 4% allele frequency. The previously reported GNAQ c.548G>A, p.Arg183Gln was confirmed in 9 of 12 cases with an allele frequency ranging from 1.73 to 7.42%. Digital polymerase chain reaction confirmed novel variants detected by next generation sequencing. Two novel somatic variants were also found in RASA1, although neither was predicted to be deleterious. Conclusions This is the second largest study on isolated, non-syndromic PWS. Our data suggest that GNAQ is the main genetic determinant in this condition. Moreover, isolated port-wine stains are distinct from capillary malformations seen in RASA1 disorders, which will be helpful in clinical evaluation. PMID:26192947

  12. Method and apparatus for staining immobilized nucleic acids

    DOEpatents

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

    2000-01-01

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

  13. New Giemsa method for the differential staining of sister chromatids

    Microsoft Academic Search

    Paul Perry; SHELDON WOLFF

    1974-01-01

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

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

    Microsoft Academic Search

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

    1985-01-01

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

  15. Peroperative fat staining of frozen sections in primary hyperparathyroidism.

    PubMed

    Ljungberg, O; Tibblin, S

    1979-06-01

    Roth and Gallaher recently described a fat staining method for rapid peroperative differentiation between parathyroid adenoma and chief cell hyperplasia. They used Sudan IV in a solution of ethanol and acetone. This solution, however, was found to cause a considerable dissolution of small lipid droplets from the tissue; in our hands sections stained with this technique were diffcult to interpret. To diminish the loss of fat from the tissue, we have used a modification of Lillie's supersaturated ispropanol method with oil red O. This method gave a deeper staining and increased the difference between hyperfunctioning and unnivolved parathyroid tissue with respect to the amount of stainable lipid in the chief cells. It was found to be a valuable supplement, adding a functional dimension to the structural interpretation of the tissue, and it facilitated the peroperative distinction between ademona and hyperplasia. The pattern of lipid distribution within the glands from patients with nodular hyperplasia suggests that the compact nodules of such glands are autonomously hyperfunctioning, whereas the intervening parts of the parenchyma are more or less responsive to the serum calcium level. PMID:88183

  16. Peroperative fat staining of frozen sections in primary hyperparathyroidism.

    PubMed Central

    Ljungberg, O.; Tibblin, S.

    1979-01-01

    Roth and Gallaher recently described a fat staining method for rapid peroperative differentiation between parathyroid adenoma and chief cell hyperplasia. They used Sudan IV in a solution of ethanol and acetone. This solution, however, was found to cause a considerable dissolution of small lipid droplets from the tissue; in our hands sections stained with this technique were diffcult to interpret. To diminish the loss of fat from the tissue, we have used a modification of Lillie's supersaturated ispropanol method with oil red O. This method gave a deeper staining and increased the difference between hyperfunctioning and unnivolved parathyroid tissue with respect to the amount of stainable lipid in the chief cells. It was found to be a valuable supplement, adding a functional dimension to the structural interpretation of the tissue, and it facilitated the peroperative distinction between ademona and hyperplasia. The pattern of lipid distribution within the glands from patients with nodular hyperplasia suggests that the compact nodules of such glands are autonomously hyperfunctioning, whereas the intervening parts of the parenchyma are more or less responsive to the serum calcium level. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:88183

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

    PubMed

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

    2012-04-30

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

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

    PubMed Central

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

    2000-01-01

    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. PMID:10788401

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

    PubMed

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

    2014-04-01

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

  20. [Laser conization guided by endocervical staining with methylene blue].

    PubMed

    Sopracordevole, F; Campagnutta, E; Parin, A; Scarabelli, C

    1994-03-01

    The authors report their experience of the use of a vital stain--methylene blue--as a surgical guide in laser cervical conization for CIN2-CIN3/CIS. During the period 1 October 1991-31 December 1992 a total of 40 laser cervical conizations were performed under local anesthesia using a CO2 laser connected to a microhandpiece and colposcope in patients with exo-endocervical lesions which were histologically positive for CIN2-CIN3/CIS. In 33/40 patients an aqueous solution of 1% methylene blue was introduced preoperatively in the endocervix using a cotton-wool bud with consequent impregnation of the pseudoglandular crypts: laser biopsy was performed along the guidelines of the stain itself. This enabled the direction of resection to be varied: in 3 patients due to an anomalous and eccentric direction of cervical canal; in 10 patients to remove glandular structures surrounding or underneath lesions; in 8 patients following pseudoglandular section to carry out deep vaporization (3 patients) or correct cutting edges (5 patients). The apex and edges of the cone were always intact. Fourteen patients completed a 12-month follow-up and a further 6 were followed up for 9 months; only 1/14 patients (with AIDS) showed recidivation after 1 year. In the authors' experience the use of a vital stain as a guide during laser cervical cone biopsy is an easily used method which ensures the greatest possible respect for healthy cervical structures, also in order to preserve fertility in young patients. PMID:8015701

  1. DAPI staining and fluorescence microscopy techniques for phytoplasmas.

    PubMed

    Andrade, Nancy M; Arismendi, Nolberto L

    2013-01-01

    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

  2. Detecting necrotic neurons with fluoro-jade stain.

    PubMed

    Krinke, G J; Classen, W; Vidotto, N; Suter, E; Würmlin, C H

    2001-10-01

    Fluoro-jade, a novel stain for detection of neuropathic lesions by fluorescence microscopy, was validated on the models of toxic neuropathy induced with 3-acetylpyridine (3-AP) or with acrylamide (ACR). Groups of male and female albino rats of Wistar strain were either exposed to a single administration of 80 mg/kg i.p. 3-AP followed 5 hours later by 300 mg/kg of nicotinamide i.p. and examined at days 3 and 15, or to 15 daily doses of 30 mg/kg p.o. ACR and examined at day 15. Following in-life behavioral observations and measurements, the rats were fixed by perfusion with formalin. Additional animals treated with same dose of 3-AP and nicotinamide were submitted to purposeful autolysis for 4 or 16 hours before immersion fixation with formalin on test day 3. In-life observations showed in 3-AP-treated animals signs of severe general toxicity, sensorimotor dysfunction and decreased motor activity starting shortly after the treatment and persisting throughout the observation period. ACR-treated rats started to develop abnormal gait on test day 8 and by day 15 developed reduced grip strength, increased landing footsplay and decreased motor activity. Fluoro-jade, applied to paraffin sections of the nervous system, detected selectively and sensitively the necrotic neurons in the brain, especially those in the inferior olivary nucleus of animals treated with 3-AP, at test day 3, as well as the necrotic Purkinje cells in the cerebellum of ACR-treated animals at test day 15. Chromatolytic neurons in the dorsal root ganglia of ACR-treated animals did not stain positively, indicating that this kind of reversible neuronal remodeling is not detectable using fluoro-jade. Necrotic neurons were still stained by fluoro-jade after 4 hour autolysis, but following 16 hour autolysis the results became false negative. There was no false positive fluorescence in fresh or autolytic tissues, except that emitted by red blood cells in unperfused specimens. The study confirmed the validity of fluoro-jade as a stain suitable for detecting necrotic neurons in toxicological safety studies. PMID:11817106

  3. Machine vision system for automated detection of stained pistachio nuts

    NASA Astrophysics Data System (ADS)

    Pearson, Tom C.

    1995-01-01

    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.

  4. Usefulness of visible dyes for the staining of protein or DNA in electrophoresis.

    PubMed

    Jin, Li-Tai; Choi, Jung-Kap

    2004-08-01

    Since 1993, we have studied visible organic dye stains for protein or DNA to improve methodologies and developed the counterion dye staining method. The method employs two oppositely charged dyes that form an ion-pair complex in the staining solution. The selective binding of free dye to protein or DNA in the staining solution improves detection sensitivity and speed. It is a rapid and sensitive procedure, involving fixing/staining or staining/quick destaining steps that are completed in 1-1.5 h. The lowest detection limits achieved are 4-8 ng of protein on polyacrylamide gels and approximately 10 ng of DNA on agarose gels. The focus of this review is to chronicle the development and current status of the counterion dye staining method for detection of protein or DNA. As an extended application of visible dyes, we also discuss the visible dye staining method for detecting protein on blotting membranes developed in our laboratory. PMID:15300759

  5. A fast silver staining method for protein detection after isoelectric focusing in agarose gels.

    PubMed

    Lasne, F; Benzerara, O; Lasne, Y

    1983-07-15

    A sensitive staining method was developed for detecting proteins in agarose gels after isoelectric focusing. Its sensitivity is about 20 times that of the Coomassie blue R-250 staining technique, and the time required is only 10 min. PMID:6194715

  6. Analysis of surface stains on modern gold coins

    NASA Astrophysics Data System (ADS)

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

    2013-07-01

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

  7. Interphase ribosomal RNA cistron staining in chronic myeloid leukaemia

    PubMed Central

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

    1995-01-01

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

  8. Vestibular schwannoma or tanycytic ependymoma: Immunohistologic staining reveals

    PubMed Central

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

    2014-01-01

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

  9. MP 33250 Coomassie FluorTM Orange Protein Gel Stain Product Information

    E-print Network

    Lebendiker, Mario

    MP 33250 Coomassie FluorTM Orange Protein Gel Stain Product Information Storage upon receipt Introduction Molecular Probes proprietary Coomassie FluorTM Orange protein gel stain provides fast, simple, and there is no risk of overstaining the gel. · Compatibility with standard laboratory equipment. Stained proteins can

  10. Performance of variations of carbolfuchsin staining of sputum smears for AFB under field conditions

    Microsoft Academic Search

    A. Van Deun; A. Hamid Salim; K. J. M. Aung; A. Hossain; N. Chambugonj; A. Hye; A. Kawria; E. Declercq

    SETTING: A field project in Bangladesh. OBJECTIVE: To compare the effectiveness of commonly used carbolfuchsin staining variations. DESIGN: Routine hot Ziehl-Neelsen (ZN) 1% basic fuchsin staining for 15 min in 75 field clinics. Blind read- ing of duplicate smears stained by ZN 1% vs. 0.3% basic fuchsin applied for 5 min, or by ZN 1% 5 min vs. Kinyoun cold

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

    E-print Network

    Løw, Erik

    LaTeX Coffee Stains Hanno Rein http://hanno-rein.de Cambridge University April 3, 2009 1 a coffee stain to your documents. A lot of time can be saved by printing stains directly on the page rather Usage To use the package, simply place the coffee.sty file in the directory with all of your other .tex

  12. A Comparison of Heat versus Methanol Fixation for Gram Staining Bacteria

    ERIC Educational Resources Information Center

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

    2009-01-01

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

  13. Use of eriochrome cyanine R for routine histology and histopathology: an improved dichromatic staining procedure.

    PubMed

    Stefanovi?, D

    2015-08-01

    A modified dichromatic iron-eriocyanine R (Fe-ECR) staining method is described. Staining obtained with this new technique generally was similar to that of hematoxylin and eosin (H & E). Cell nuclei were stained blue. Cardiac, smooth and skeletal muscle, and red blood cells, were stained different shades of red. Collagen fibers were stained different shades of orange, usually faintly. Decalcified bony tissue was stained pinkish violet. Epithelial cells were strongly stained deep shades of red, magenta and violet. Cartilage matrix, and goblet and mast cells were unstained. Although Fe-ECR staining differed too much from standard H & E staining to be a substitute for diagnostic purposes, the dichromatic method described might usefully replace van Gieson or trichrome stains, especially if muscle is of interest. A pH 0.95 staining solution was used to differentiate initially over-stained sections followed by washing in distilled water. This dichromatic technique is easier to perform and more precisely controllable than other ECR dichromatic methods. The entire procedure can be completed in less than 5 min. The technique has the advantages of greater technical simplicity and speed, a larger range of polychromasia, and a longer shelf-life than H & E. ECR also is more reliably available than hematoxylin and usually is less expensive. PMID:26140653

  14. FACS Staining Protocol for hair follicle stem cells Elizabeth Deschene Greco Lab

    E-print Network

    Greco, Valentina

    FACS Staining Protocol for hair follicle stem cells Elizabeth Deschene ­ Greco Lab Color compensation controls are samples that are stained with only one of the color fluorophores sample. If you are staining for hair germ and bulge cells and when using Aria (for sorting) or LSRII

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

    Microsoft Academic Search

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

    1996-01-01

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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  17. 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

    Ankle, Madhuri R; Joshi, Priya S

    2011-01-01

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

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

    PubMed

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

    2014-10-01

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

  19. Effects of activated macrophages on Mycobacterium leprae.

    PubMed Central

    Ramasesh, N; Adams, L B; Franzblau, S G; Krahenbuhl, J L

    1991-01-01

    Five alternative methods were used to explore in vitro the effects of normal and activated murine macrophages on the metabolic well-being of intracellular Mycobacterium leprae: fluorescein diacetate-ethidium bromide staining, ATP content, synthesis of phenolic glycolipid 1, and two techniques to quantitate oxidation of palmitic acid. In relatively short-term experiments (7 to 10 days), each of these procedures provided strong evidence that activated macrophages exerted a deleterious effect on the leprosy bacillus. These findings appear to confirm the contention that activated macrophages underlie host resistance to clinical leprosy and limitation of M. leprae growth in paucibacillary leprosy. PMID:1908824

  20. Fat tissue staining and photodynamic/photothermal effects

    NASA Astrophysics Data System (ADS)

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

    2010-02-01

    Cellulite is considered as a disease of the subcutaneous fat layer that appears mostly in women and consists of changes in fat cell accumulation together with disturbed lymphatic drainage, affecting the external appearance of the skin. The photodynamic and selective photothermal treatments may provide reduction the volume of regional or sitespecific accumulations of subcutaneous adipose tissue on the cellular level. We hypothesize that light irradiation of stained fat tissue at selected temperature leads to fat cell lypolytic activity (the enhancement of lipolysis of cell triglycerides due to expression of lipase activity and cell release of free fat acids (FFAs) due to temporal cell membrane porosity), and cell killing due to apoptosis caused by the induced fat cell stress and/or limited cell necrosis.

  1. Discoloration of coating resins exposed to staining solutions in vitro.

    PubMed

    Manabe, Atsufumi; Kato, Yukiyo; Finger, Werner J; Kanehira, Masafumi; Komatsu, Masashi

    2009-05-01

    The purpose of this in vitro investigation was to study the effects of coffee and tea immersion on surface discoloration of one commercial temporary resin coating material, White Coat (WHC; Kuraray Medical Inc., Tokyo, Japan), and an experimental one, SI-R20209 (SIR; Shofu Inc., Kyoto, Japan). Disk-shaped specimens were prepared, their colors were determined at baseline, and after immersion in water (control), tea and coffee solutions for 24, 48, and 72 hours. Very little discoloration was found with the water-stored specimens. Staining response was most pronounced after coffee immersion for White Coat and after tea immersion for the experimental material, exceeding the clinically acceptable discoloration threshold value of deltaE=3.3. However, most of the resin shades tested are likely to be sufficiently safe against heavy discoloration when used for short-term restoration only. PMID:19662733

  2. Early morphea mimicking acquired port-wine stain.

    PubMed

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

    2014-01-01

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

  3. Selective gray matter staining of human brain slices: optimized use of cadaver materials.

    PubMed

    Loftspring, M C; Smanik, J; Gardner, C; Pixley, S K

    2008-06-01

    We report a novel staining technique for human brain slices that distinguishes clearly gray from white matter. Previously described techniques using either Prussian blue (Berlin blue) or phthalocyanine dyes usually have included a hot phenol pretreatment to prevent white matter staining. The technique we describe here does not require hot phenol pretreatment and allows the use of brains stored for postmortem periods of one to two years prior to staining. Our technique involves staining with copper(II) phthalocyanine-tetrasulfonic acid tetrasodium salt 1% in water for 2 h followed by acetic acid treatment; this produces excellent blue staining of gray matter with little white matter staining. The stained brain slices are excellent for teaching human brain anatomy and/or pathology, or for research purposes. PMID:18946763

  4. Multi-stained whole slide image alignment in digital pathology

    NASA Astrophysics Data System (ADS)

    Déniz, Oscar; Toomey, David; Conway, Catherine; Bueno, Gloria

    2015-03-01

    In Digital Pathology, one of the most simple and yet most useful feature is the ability to view serial sections of tissue simultaneously on a computer monitor. This enables the pathologist to evaluate the histology and expression of multiple markers for a patient in a single review. However, the rate limiting step in this process is the time taken for the pathologist to open each individual image, align the sections within the viewer, with a maximum of four slides at a time, and then manually move around the section. In addition, due to tissue processing and pre-analytical steps, sections with different stains have non-linear variations between the two acquisitions, that is, they will stretch and change shape from section to section. To date, no solution has come close to a workable solution to automatically align the serial sections into one composite image. This research work address this problem to obtain an automated serial section alignment tool enabling the pathologists to simply scroll through the various sections in a single viewer. To this aim a multi-resolution intensity-based registration method using mutual information as a similarity metric, an optimizer based on an evolutionary process and a bilinear transformation has been used. To characterize the performance of the algorithm 40 cases x 5 different serial sections stained with hematoxiline-eosine (HE), estrogen receptor (ER), progesterone receptor (PR), Ki67 and human epidermal growth factor receptor 2 (Her2), have been considered. The qualitative results obtained are promising, with average computation time of 26.4s for up to 14660x5799 images running interpreted code.

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

    PubMed

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

    2014-08-01

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

  6. Staining of in vivo subsurface degradation in dental composites with silver nitrate

    SciTech Connect

    Mair, L.H. (Univ. of Liverpool (England))

    1991-03-01

    A previously reported technique for staining areas of degradation in dental composite restorations was evaluated in 51 removed restorations. The staining reagent was silver nitrate, which penetrated the degraded subsurface as ionic silver and was subsequently developed into colored deposits of metallic silver. Several artefacts were recognized that resulted in an apparent image of subsurface stain. Most importantly, the presence of a layer of adsorbed silver on the edge of the specimen exaggerated the extent of staining. In order for the true depth of stain to be determined, thin sections of the materials should first be examined with a stereomicroscope to distinguish any contribution from adsorbed silver on the specimen edge. With this regimen, no stain was present in 41% of the restorations, and in a further 30%, the depth of stain was less than 50 microns. In two composites, the depth of stain was greater than 900 microns, and in a number of specimens, localized stain was found in association with attrition scars. Energy-dispersive x-ray analysis indicated that the amount of silver present in the degraded layers was very small. Overall, the results indicated that the staining technique is useful in the study of composite degradation.

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

    PubMed

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

    2012-11-30

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

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

    PubMed

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

    2015-02-01

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

  9. Effect of age of cook-in-bag delicatessen meats formulated with lactate-diacetate on the behavior of Listeria monocytogenes contamination introduced when opening the packages during storage.

    PubMed

    Geornaras, Ifigenia; Toczko, Darren; Sofos, John N

    2013-07-01

    This study evaluated the potential effect of age of cook-in-bag ham and turkey breast delicatessen meats formulated with lactate-diacetate on survival and/or growth of Listeria monocytogenes introduced after opening of packages and slicing of product. Commercially prepared cured ham and turkey breast products formulated with potassium lactate and sodium diacetate were stored at 1.7°C unsliced, in their original cook-in-bags, and without postlethality exposure. On days 5, 90, 120, and 180 of storage, product slices (10.2 by 7.6 cm) were surface inoculated (1 to 2 log CFU/cm²) with a 10-strain mixture of L. monocytogenes, vacuum packaged (seven slices per bag), and stored at 4°C for up to 13 weeks. Inoculated levels of L. monocytogenes on both products were 1.4 to 1.5 log CFU/cm². Irrespective of product age at slicing and inoculation, after 13 weeks of vacuum-packaged storage (4°C), pathogen counts on product slices were 1.5 to 2.3 (ham) and 2.3 to 2.5 (turkey) log CFU/cm². Overall, the results of the study showed that the age of the cook-in-bag products prior to slicing and inoculation with the pathogen did not (P ? 0.05) affect the behavior of L. monocytogenes during vacuum-packaged storage (4°C, up to 13 weeks) of ham and turkey slices. Mean counts of lactic acid bacteria and yeasts and molds, when detected, did not exceed approximately 1 and 2 log CFU/cm², respectively, among all stored samples. Findings of the study will be useful to the meat industry and risk assessors in their efforts to control L. monocytogenes in ready-to-eat meat products. PMID:23834806

  10. Pneumocystis carinii pneumonia in patients with AIDS: evaluation of lavage and staining techniques in diagnosis.

    PubMed

    Chechani, V; Allam, A A; Haseeb, M A; Kamholz, S L

    1991-01-01

    The diagnostic yield of unilateral vs. bilateral bronchoalveolar lavage (BAL) was prospectively evaluated in 65 consecutive patients suspected of having Pneumocystis carinii pneumonia (PCP) complicating acquired immune deficiency syndrome (AIDS). Gram-Weigert (GW), Papanicolaou (PAP), and Gomori's methenamine silver (GMS) stains were used for identification of P. carinii in all cases. Forty-eight patients had PCP that was identified by GW staining of BAL in 47/48 patients followed by PAP/GMS staining of BAL in 44/48 patients and PAP/GMS staining of bronchial washings in 40/48 patients. In patients with bilateral interstitial infiltrates, unilateral lavage was sufficient for diagnosis of PCP when GW stain was utilized. In patients with PCP complicating AIDS, the diagnostic yield of BAL may be increased by use of both GW and GMS stains. PMID:1704061

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

    PubMed

    Farrugia, Kevin J; Savage, Kathleen A; Bandey, Helen; Nic Daéid, Niamh

    2011-09-01

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

  12. Fluorescent staining for leukocyte chemotaxis. Eosinophil-specific fluorescence with aniline blue.

    PubMed

    McCrone, E L; Lucey, D R; Weller, P F

    1988-11-10

    To overcome problems associated with the quantitation of human eosinophil chemotaxis in micropore filters, we have developed a fluorescent method of specifically staining eosinophils in chemotactic filters. A neutral solution of aniline blue yielded bright green fluorescent staining of the cytoplasmic granules of eosinophils. Other leukocytes and contaminating neutrophils potentially present with eosinophils did not fluoresce with aniline blue. The fluorescent staining eosinophils within filters provided bright, non-fading images that facilitated visual microscopic counting and were of sufficiently high contrast, unlike those with conventional eosinophil stains, to allow image analyzer based enumeration of eosinophil chemotactic responses at levels through the filters. Although not cell type-specific, congo red and ethidium bromide also provided high contrast, fluorescent images of all leukocyte types within chemotactic filters. Fluorescent staining with aniline blue constitutes a rapid, stable and eosinophil-specific stain that facilitates the visual or image analyzer-based quantitation of eosinophil chemotaxis. PMID:2460564

  13. Diffuse reflectance FTIR of stains on grit blasted metals

    SciTech Connect

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

    1997-08-09

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

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

    PubMed

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

    2014-12-01

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

  15. Isolation of Escherichia coli mannitol permease, EIImtl, trapped in amphipol A8-35 and fluorescein-labeled A8-35.

    PubMed

    Opa?i?, Milena; Giusti, Fabrice; Popot, Jean-Luc; Broos, Jaap

    2014-10-01

    Amphipols (APols) are short amphipathic polymers that keep integral membrane proteins water-soluble while stabilizing them as compared to detergent solutions. In the present work, we have carried out functional and structural studies of a membrane transporter that had not been characterized in APol-trapped form yet, namely EII(mtl), a dimeric mannitol permease from the inner membrane of Escherichia coli. A tryptophan-less and dozens of single-tryptophan (Trp) mutants of this transporter are available, making it possible to study the environment of specific locations in the protein. With few exceptions, the single-Trp mutants show a high mannitol-phosphorylation activity when in membranes, but, as variance with wild-type EII(mtl), some of them lose most of their activity upon solubilization by neutral (PEG- or maltoside-based) detergents. Here, we present a protocol to isolate these detergent-sensitive mutants in active form using APol A8-35. Trapping with A8-35 keeps EII(mtl) soluble and functional in the absence of detergent. The specific phosphorylation activity of an APol-trapped Trp-less EII(mtl) mutant was found to be ~3× higher than the activity of the same protein in dodecylmaltoside. The preparations are suitable both for functional and for fluorescence spectroscopy studies. A fluorescein-labeled version of A8-35 has been synthesized and characterized. Exploratory studies were conducted to examine the environment of specific Trp locations in the transmembrane domain of EII(mtl) using Trp fluorescence quenching by water-soluble quenchers and by the fluorescein-labeled APol. This approach has the potential to provide information on the transmembrane topology of MPs. PMID:24952466

  16. Distamycin A\\/DAPI staining of heterochromatin in male meiosis of man

    Microsoft Academic Search

    T. Haaf; H. Miiller; M. Schmid

    1986-01-01

    The sequential staining with distamycin A\\/DAPI provides an ideal method for studying the behaviour of heterochromatic regions in human male meiosis. The various meiotic and postmeiotic stages were found to have different staining qualities. Although all heterochromatic regions in human pachytene cells show specific DA\\/DAPI fluorescence, bright and clearly stained heterochromatic blocks can be distinguished from small DA\\/DAPI spots. Pachytene

  17. Juvenile localized scleroderma with port wine stain: coincidental or possible common pathogenetic association.

    PubMed

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

    2015-01-01

    Port wine stain and juvenile localized scleroderma are two different dermatoses usually encountered in pediatric age group. Up to now, there are reports of morphea patients initially diagnosed and treated as port wine stain. Coexistence of both diseases is not found yet. We herein present a case of juvenile localized scleroderma on the left side of trunk, with congenital port wine stain located on the ipsilateral face at V1-V2 distribution. PMID:25814757

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

    PubMed Central

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

    2015-01-01

    Port wine stain and juvenile localized scleroderma are two different dermatoses usually encountered in pediatric age group. Up to now, there are reports of morphea patients initially diagnosed and treated as port wine stain. Coexistence of both diseases is not found yet. We herein present a case of juvenile localized scleroderma on the left side of trunk, with congenital port wine stain located on the ipsilateral face at V1-V2 distribution. PMID:25814757

  19. Multispectral image enhancement for H&E stained pathological tissue specimens

    NASA Astrophysics Data System (ADS)

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

    2008-03-01

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

  20. Stevenel's Blue, an excellent stain for optical microscopical study of plastic embedded tissues.

    PubMed

    del Cerro, M; Cogen, J; del Cerro, C

    1980-05-01

    Stevenel's Blue is a reliable, rapid, and clean, one-step polychromatic stain for 1 micron thick epoxy sections. The staining solution, originally used by L. Stevenel (1918) to stain human parasites, is made by adding diluted potassium permanganate (2%) to an aqueous solution of the methylene blue (1.3%) and redissolving the precipitate thoroughly, by boiling in water bath and filtering. Staining is carried out in a Coplin jar at 60 degrees C for approximately 10 minutes for tissues embedded in Epon 812 or Poly 812, or 20 minutes for tissues embedded in Spurr's medium. The sections are rinsed, air dried, and mouted in Permount. The staining solution is very stable, and does not tend to form precipitates on the tissue. The stain brings excellent histological differentiation to nuclear, cytoplasmic, and extracellular components. Incorporation of the stain by elements within each tissue varies from intense to light with a subtle gradation of intermediate shades of purple and blue tones. For most cell structures the density of the stain parallels the electron density of that structure as seen under the electron microscope. For example, nucleoli and heterochromatin stain in dark purple while euterochromatin appear in a light blue shade. In all cases, the embedding media remains unstained. The bond between Stevenel's Blue and the tissues is stable, remaining unaltered by the mounting medium. It is also resistant to time-fading. PMID:6156384

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

    PubMed Central

    Karagyan, Gayane; Lachowska, Dorota; Kalashian, Mark

    2012-01-01

    Abstract The male karyotypes of Acmaeodera pilosellae persica Mannerheim, 1837 with 2n=20 (18+neoXY), Sphenoptera scovitzii Faldermann, 1835 (2n=38–46), Dicerca aenea validiuscula Semenov, 1895 – 2n=20 (18+Xyp) and Sphaerobothris aghababiani Volkovitsh et Kalashian, 1998 – 2n=16 (14+Xyp) were studied using conventional staining and different chromosome banding techniques: C-banding, AgNOR-banding, as well as fluorochrome Chromomycin A3 (CMA3) and DAPI. It is shown that C-positive segments are weakly visible in all four species which indicates a small amount of constitutive heterochromatin (CH). There were no signals after DAPI staining and some positive signals were discovered using CMA3 staining demonstrating absence of AT-rich DNA and presence of GC-rich clusters of CH. Nucleolus organizing regions (NORs) were revealed using Ag-NOR technique; argentophilic material mostly coincides with positive signals obtained using CMA3 staining. PMID:24260661

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

    Microsoft Academic Search

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

    1997-01-01

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

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

    PubMed

    Ousler, George; Devries, Douglas K; Karpecki, Paul M; Ciolino, Joseph B

    2015-01-01

    A single-center, open-label study consisting of two visits over the course of approximately 2 weeks was conducted to evaluate the efficacy of Retaine™ ophthalmic emulsion in improving the signs and symptoms of dry eye. Forty-two subjects were enrolled and received 1-2 drops twice daily of Retaine™ beginning at the first visit (day 1) and ending at the second visit. Subjects were instructed to complete a symptomatology diary twice daily prior to drop instillation through the morning of the second visit. Ocular sign and symptom assessments, visual acuity procedures, and comfort assessments were conducted during both visits. A statistically significant reduction was observed in mean breakup area on the second visit between the predose time and the postdose time (P=0.026). On the second visit, subjects had significantly less corneal fluorescein staining in the superior (P=0.002), central (P=0.017), corneal sum (P=0.011), and all ocular regions combined (P=0.038) than on the first visit. On the second visit, statistically significant reductions in dryness (P<0.001), grittiness (P=0.0217), ocular discomfort (P=0.0017), and all symptoms (P<0.001) were also seen as measured by the Ora Calibra™ Ocular Discomfort and 4-Symptom Questionnaire (0-5 scale). Subjects reported a statistically significant improvement in their abilities to work with a computer at night (P=0.044). Mean drop comfort scores ranged from 1.29-1.81 on the Ora Calibra™ 0-10 Drop Comfort Scale, on which 0 is very comfortable and 10 is very uncomfortable. Retaine™ demonstrates promising results as a novel artificial tear option for individuals suffering from dry eye. The unique mechanism of action of Retaine™ provides enhanced comfort and improves the quality of life of dry eye subjects while reducing the ocular signs of dry eye. PMID:25709384

  4. Energy transfer and binding competition between dyes used to enhance staining differentiation in metaphase chromosomes

    Microsoft Academic Search

    Elhanan Sahar; Samuel A. Latt

    1980-01-01

    The ability of electronic energy transfer and direct binding competition between pairs of dyes to enhance contrast in human or bovine metaphase chromosome staining patterns is illustrated, and the relative effectiveness of these two mechanisms compared. The existence of energy transfer between quinacrine or 33258 Hoechst and 7-amino-actinomycin D in doubly stained chromosomes is demonstrated directly by microfluorometry. The ability

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

    Microsoft Academic Search

    MICHAEL L. WATSON; WILLIAM G. ALDRIDGE

    1961-01-01

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

  6. Little Strokes Fill Big Oaks: A Simple In Vivo Stain of Brain Cells

    E-print Network

    Bern, Universität

    Neuron Previews Little Strokes Fill Big Oaks: A Simple In Vivo Stain of Brain Cells Fritjof Helmchen1, * and Thomas Nevian2 1 Department of Neurophysiology, Brain Research Institute, University, ``The gain in brain is mainly in the stain.'' This notion clearly ex- presses the fundamental importance

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

    SciTech Connect

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

    2002-05-01

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

  8. A C. elegans mutant screen based on antibody or histochemical staining

    Microsoft Academic Search

    Guofeng Xie; Yiwen Jia; Eric Aamodt

    1995-01-01

    A method has been developed for isolating mutations in Caenorhabditis elegans that alter antibody or histochemical staining patterns. The basis for this method is a new procedure for making C. elegans permeable that does not kill the eggs contained within the uterus of gravid adult hermaphrodites. A mutagenized population of gravid hermaphrodites is made permeable and then stained with either

  9. Effect of porous silicon stain etched on large area alkaline textured crystalline silicon solar cells

    Microsoft Academic Search

    N. Marrero; R. Guerrero-Lemus; B. González-Díaz; D. Borchert

    2009-01-01

    This work shows the effects of porous silicon stain etched on alkaline textured antireflection coatings of large area monocrystalline silicon solar cells. The texturization process has been produced by immersion of the silicon wafers in different carbonate-based solutions. The porous silicon layers were formed by stain etching in a HNO3\\/HF aqueous solution before or after the texturization process. We study

  10. 9 CFR 147.3 - The stained-antigen, rapid, whole-blood test. 3

    Code of Federal Regulations, 2012 CFR

    2012-01-01

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

  11. 9 CFR 147.3 - The stained-antigen, rapid, whole-blood test. 3

    Code of Federal Regulations, 2014 CFR

    2014-01-01

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

  12. 9 CFR 147.3 - The stained-antigen, rapid, whole-blood test. 3

    Code of Federal Regulations, 2013 CFR

    2013-01-01

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

  13. Postharvest changes in pigment concentrations in ‘Fuji’ apples with ‘Fuji’ stain

    Microsoft Academic Search

    David A. Felicetti; Larry E. Schrader

    2010-01-01

    ’Fuji’ apples (Malus domestica Borkh cv. ‘Fuji’) sometimes develop a discolouration in the peel during cold storage, typically in the periphery of sunburned peel. We refer to this particular postharvest disorder as ‘Fuji’ stain as we have not observed it in any cultivar other than ‘Fuji’ and the discolouration looks like a stain on the peel. Because peel discolouration occurs,

  14. Staining correction in digital pathology by utilizing a dye amount table.

    PubMed

    Bautista, Pinky A; Yagi, Yukako

    2015-06-01

    The stained colors of the tissue components are popularly used as features for image analysis. However, variations in the staining condition of the histology slides prompt variations to the color distribution of the stained tissue samples which could impact the accuracy of the analysis. In this paper, we present a method to correct the staining condition of a histology image. In the method, a look-up table (LUT) based on the dye amounts absorbed by the sample is built. The LUT can be built when either (i) the source and reference staining conditions are specified or (ii) when the user simply wants to recreate his/her preferred staining condition without specifying any reference slide. The effectiveness of the present method was evaluated in two aspects: (i) CIELAB color difference of nuclei, cytoplasm, and red blood cells, between the ten different slides of liver tissue, and (ii) classification of the different tissue components. Application of the present staining correction method reduced the color difference between the slides by an average factor of 9.8 and the classification performance of a linear discriminant classifier improved by 16.5 % on the average. Results of the paired t test statistical analysis further showed that the reduction in the CIELAB color difference between the slides and the improvement in the classifier's performance when staining correction was implemented is significant at p?

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

    E-print Network

    Paris-Sud XI, Université de

    Specific In Vivo Staining of Astrocytes in the Whole Brain after Intravenous Injection In Vivo Staining of Astrocytes in the Whole Brain after Intravenous Injection of Sulforhodamine Dyes. PLo brain [10,11,12] and has opened a new field of dynamic and functional studies on neuron-astrocytes

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  17. EDTA Separation and ATPase Langerhans Cell Staining in the Mouse Epidermis

    Microsoft Academic Search

    Kenneth W. Baker; Joseph E. J. Habowsky

    1983-01-01

    Sheets of ethylenediaminetetraacetic acid (EDTA)- separated epidermis were examined using scanning electron, transmission electron, and light microscopy; sheets were also examined after staining for adenosine triphosphatase (ATPase) activity. Staining was improved by longer incubation with EDTA and by elimination of Trismal buffer as a tissue rinse. EDTA-separated epidermis showed better retention of ultrastructural integrity when washed with phosphate-buffered saline. The

  18. Sodium thiosulfate solution spray for relief of irritation caused by Lugol's stain in chromoendoscopy

    Microsoft Academic Search

    Hitoshi Kondo; Haruhiko Fukuda; Hiroyuki Ono; Takuji Gotoda; Daizo Saito; Kozu Takahiro; Kuniaki Shirao; Hajime Yamaguchi; Shigeaki Yoshida

    2001-01-01

    Background: Mucosal iodine staining is known to improve the endoscopic visualization of esophageal squamous dysplasia and cancer. However, it often causes mucosal irritation leading to retrosternal discomfort. The clinical usefulness of sodium thiosulfate solution (STS) for easing symptoms induced by mucosal staining with Lugol's solution was evaluated in this study. Methods: One hundred twenty healthy men over 50 years of

  19. Haemolymph flows in the wings of pierid butterflies visualized by vital staining (insecta, lepidoptera)

    Microsoft Academic Search

    Lutz Thilo Wasserthal

    1983-01-01

    The flow of stained haemolymph was photographed in the wings of resting Pieris rapae, Pieris brassicae, and Gonepteryx rhamni under UV-radiation at definite intervals after abdominal application of fluorescent tetracycline. There is no circular route in the wing. All wing veins are supplied with stained haemolymph from their own bases without preference to single veins. In freely resting Pieris with

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

    PubMed

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

    2013-06-25

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

  1. When Conjugated Polymers Meet Amyloid Cliff I. Stains and Indraneel Ghosh*

    E-print Network

    Ghosh, Indraneel

    along with neurofibrillary tangles that stain with Congo red in the brains of patients. The A peptides of A ) in the brain and early cogni- tive dysfunction than between the A de- posits stained by Congo red that soluble oligomeric intermediates rather than the final fibrils are responsible for neu- rological toxicity

  2. Vital staining of specific monoamine-containing cells in the leech nervous system

    Microsoft Academic Search

    Ann E. Stuart; A. J. Hudspeth; Zach W. Hall

    1974-01-01

    Neutral red and several related dyes selectively stain certain cells in the ventral nerve cord of the leech. These cells are identical with those that can be shown by the FalckHillarp fluorescence technique to contain serotonin or a catecholamine; evidence suggests that the catecholamine is dopamine. Although the mechanism of staining remains unknown, it does not depend on active uptake

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

    Code of Federal Regulations, 2011 CFR

    2011-01-01

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

  4. An improved silver staining technique for nucleolus organizer regions by using nylon cloth

    Microsoft Academic Search

    Yoshiaki Kodama; Michihiro C. Yoshida; Motomichi Sasaki

    1980-01-01

    Summary A simple and reproducible silver-staining technique for nucleolus organizer regions (NORs) was developed, use being made of nylon cloth as a coverslip for even impregnation of the silver solution. Ag-NORs were clearly and selectively visualized in human and mouse chromosomes, without equivocal staining of centrometric heterochromatin and background silver grains.

  5. Using stained glass windows to understand the durability of toxic waste matrices

    Microsoft Academic Search

    Jérôme Sterpenich; Guy Libourel

    2001-01-01

    Using stained glasses sampled from French and German cathedrals, and from different archaeological sites, this work presents an estimation of the effect of weathering conditions and glass composition on glass dissolution. Due to accurate dating, we also show that stained glass windows allow the determination of the average dissolution kinetics of many toxic elements contained in the glass (including transition

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

    E-print Network

    Paris-Sud XI, Université de

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

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

    NASA Astrophysics Data System (ADS)

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

    2011-10-01

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

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

    PubMed Central

    Clarke, S C; McIntyre, M

    1996-01-01

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

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

    PubMed

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

    2000-01-01

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

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

    PubMed

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

    2014-12-01

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

  11. Differentiating Taenia eggs found in human stools - Does Ziehl Neelsen staining help?

    PubMed Central

    Jimenez, Juan A.; Rodriguez, Silvia; Moyano, Luz M.; Castillo, Yesenia; García, Héctor H.

    2012-01-01

    SUMMARY Introduction Unlike other tapeworms, T. solium infections carry risk for neurocysticercosis. Differential diagnosis of human tapeworm infections relies on morphology of the scolex or proglottids, frequently unavailable. DNA-based assays are poorly available in endemic areas. Ziehl Neelsen staining has been suggested but not tested in controlled designs. We validated whether Ziehl Neelsen staining could differentiate T. solium and T. saginata eggs. Methods Tapeworm proglottids (33 specimens, 23 T. solium and 10 T. saginata) and eggs (31 specimens, 13 T. solium and 10 T. saginata) were stained. Four eggs from each sample were measured and average diameters were recorded. Results T. saginata eggs stained entirely magenta in seven of 13 cases. T. solium eggs stained entirely blue/purple in 4/18 cases and entirely magenta in one. Eggs of T. saginata were slightly larger and always ovoid, while T. solium eggs were smaller and were mostly spheric. Conclusions Ziehl Neelsen staining can occasionally distinguish fully mature T. solium from T. saginata eggs. This distinction is poorly sensitive and not completely specific. Differential staining suggest differences in embryophore components between species, evident along egg maturation. In this small series, egg morphology (shape, maximal diameter) provided appropriate differentiation between T. solium and T. saginata eggs. PMID:20579318

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

    NASA Technical Reports Server (NTRS)

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

    1991-01-01

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

  13. Abelardo Gallego (1879-1930) and his contributions to histotechnology: the Gallego stains.

    PubMed

    Ortiz-Hidalgo, Carlos

    2011-02-01

    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 enables a complete differentiation between elastic and collagenous connective tissue. The elastic tissue is stained in brilliant fuschin red, while collagen is stained in brilliant green. Abelardo Gallego was born in Spain on September 10, 1879. He was appointed as professor of Pharmacology and Therapeutics at the Faculty of Veterinary Medicine at the University of Santiago de Compostela, Spain, and in 1921 became full-time professor of Histology and Pathology in Madrid supported by Ramón y Cajal. Gallego conducted extensive research into veterinary pathology. Don Abelardo was a quiet, reserved, but friendly, sensitive, optimistic and persistent person. Weakened by a bout of bronchitis, Gallego died aged 51 on February 3, 1930, and was buried at the Almudena Cemetery, in Madrid. Gallego was one of the most outstanding scientists of Spanish veterinary medicine and some of his achievements are reviewed. PMID:19853895

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

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2014-07-01

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

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

    PubMed

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

    2012-07-10

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

  17. An adaptive algorithm for detection of multiple-type, positively stained nuclei in IHC images with minimal prior information: application to OLIG2 staining gliomas

    NASA Astrophysics Data System (ADS)

    Akakin, Hatice C.; Gokozan, Hamza; Otero, Jose; Gurcan, Metin N.

    2015-03-01

    We propose a method to detect and segment the oligodendrocytes and gliomas in OLIG2 immunoperoxidase stained tissue sections. Segmentation of cell nuclei is essential for automatic, fast, accurate and consistent analysis of pathology images. In general, glioma cells and oligodendrocytes mostly differ in shape and size within the tissue slide. In OLIG2 stained tissue images, gliomas are represented with irregularly shaped nuclei with varying sizes and brown shades. On the other hand, oligodendrocytes have more regular round nuclei shapes and are smaller in size when compared to glioma cells found in oligodendroglioma, astrocytomas, or oligoastrocytomas. The first task is to detect the OLIG2 positive cell regions within a region of interest image selected from a whole slide. The second task is to segment each cell nucleus and count the number of cell nuclei. However, the cell nuclei belonging to glioma cases have particularly irregular nuclei shapes and form cell clusters by touching or overlapping with each other. In addition to this clustered structure, the shading of the brown stain and the texture of the nuclei differ slightly within a tissue image. The final step of the algorithm is to classify glioma cells versus oligodendrocytes. Our method starts with color segmentation to detect positively stained cells followed by the classification of single individual cells and cell clusters by K-means clustering. Detected cell clusters are segmented with the H-minima based watershed algorithm. The novel aspects of our work are: 1) the detection and segmentation of multiple-type, positively-stained nuclei by incorporating only minimal prior information; and 2) adaptively determining clustering parameters to adjust to the natural variation in staining as well as the underlying cellular structure while accommodating multiple cell types in the image. Performance of the algorithm to detect individual cells is evaluated by sensitivity and precision metrics. Promising segmentation results (91% sensitivity and 86% precision) were achieved for a dataset of fourteen tissue slides with ground truth markings by two pathologists.

  18. Surface and statistical analysis of CaCO 3 crystals synthesized in the presence of fluorescein-tagged starch grafted with polyacrylic acid

    NASA Astrophysics Data System (ADS)

    Matahwa, H.; Sanderson, R. D.

    2009-02-01

    Crystallization of CaCO 3 was carried out in the presence of fluorescein-tagged starch grafted with polyacrylic acid (PAA) as polymeric additive. The crystal morphology at high polymeric additive concentration was spherical, with vaterite and calcite polymorphs. Mixed crystal morphologies were obtained at lower concentration of the polymeric additive and calcite was obtained. The CaCO 3 crystals obtained fluoresced under UV irradiation showing the presence of grafted starch expected on the surface of CaCO 3. The zeta potentials of the synthesized crystals were negative and addition of cationic starch lead to inversion of the zeta potential. Rodamine B-tagged cationic starch was successfully deposited onto the surface of the anionic-grafted starch-coated CaCO 3 crystals. Statistical analysis of the crystals showed that the fluorescence intensities of the crystals increased with increasing granularity and size of the CaCO 3 crystals. The high granularity and large-size crystals were due to aggregation of secondary crystals as observed with fluorescence microscopy.

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

    NASA Astrophysics Data System (ADS)

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

    2013-11-01

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

  20. Effect of Staining Solutions on Color Stability of Silorane & Methacrylate Restorative Material

    PubMed Central

    S. Madhyastha, Prashanthi; G. Naik, Dilip; Kotian, Ravindra; Srikant, N.; M. R. Bhat, Kumar

    2015-01-01

    Color stability throughout the functional lifetime of restorations is important for the durability of treatment and of cosmetic importance. The purpose of this study was to evaluate the discoloration properties of a silorane-based (Filtek P90) and methacrylate-based (Z100) composites upon exposure to different staining solutions that are used on day to day basis (turmeric, tea, coffee, cocoa, lime, yoghurt and distilled water) for different immersion periods (1, 7, 14 and 28 days). The colors of all specimens before and after storage in the solutions were measured by a reflectance spectrophotometer based on CIE Lab system and the color differences were calculated. Data were statistically analyzed by repeated measures of ANOVA and sidak post hoc test (for immersion period);‘t’ test (for each material) and one way ANOVA (for staining agents). All the staining agents showed significant difference in staining over time in both the materials. However, Z100 showed higher quantum of discoloration at all time periods at each staining agents (p<0.005). In conclusion, the silorane-based resin (Filtek P90) composites exhibited better color stability (less change in ?E) after exposure to the staining solutions. Among the staining agents cocoa was found to be least staining followed by lime, yoghurt, coffee, tea whereas turmeric discolored the composites to the maximum. Highest discoloration was seen at day 28 in all staining agents. Cocoa and lime discolored to maximum at early stages but remained stable thereafter whereas tea, coffee and turmeric progressively discolored the composite over time.

  1. Intracellular cytokine detection by flow cytometry in pigs: fixation, permeabilization and cell surface staining.

    PubMed

    Zelnickova, Petra; Faldyna, Martin; Stepanova, Hana; Ondracek, Jaroslav; Kovaru, Frantisek

    2007-10-31

    Intracellular flow cytometry is a method of cytokine detection that allows simultaneous detection of intracellular cytokines and cell surface markers. This important method is not extensively used in pigs, in particular due to the inaccessibility of proper methodological protocols modifying comprehensive human protocols. The aim of this study was to find the best procedure for fixation and permeabilization of porcine blood leukocytes and simultaneous cell surface staining. Permeabilization with commercial kits gave better results in most of the chosen parameters compared with combinations of different concentrations of paraformaldehyde and saponin. Among the commercial kits tested, the best results were obtained with the IntraStain kit. Cell surface markers were detected on cells stimulated for cytokine production by antibodies anti-CD14 (clone MIL-2), anti-SWC3, anti-CD4 and anti-CD8 except anti-CD14 (clone Tük4). While anti-CD8 (clone MIL-12) must be used for staining of unfixed cells, the other antibodies recognize fixed and/or permeabilized cells. Moreover, anti-SWC3 and anti-CD14 (clone MIL-2) antibodies can stain cells during the permeabilization step. These modifications of the cell surface staining protocol allow the researcher to speed up the procedure of intracellular cytokine staining or to combine cell surface staining and intracellular cytokine staining. The present study can serve as a particular protocol of intracellular cytokine detection and as a suggestion for optimization of the fixation, permeabilization and cell surface staining procedure in any laboratory. PMID:17720184

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

    NASA Technical Reports Server (NTRS)

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

    1999-01-01

    Exposure to microgravity during space flight results in profound physiologic changes. Numerous studies have shown changes in circulating populations of peripheral blood immune cells immediately after space flight. It is currently unknown if these changes result from exposure to microgravity or are caused by the stress of reentry and readaptation to gravity. We have developed the whole blood staining device as a system for the staining of whole blood collected during space flight for subsequent flow cytometric analysis, This device contains all liquids to address safety issues concerned with space flight and also moves the cells through the staining, lyse/fixation and dilution steps.

  3. An indoxyl compound 5-bromo-4-chloro-3-indolyl 1,3-diacetate, CAC-0982, suppresses activation of Fyn kinase in mast cells and IgE-mediated allergic responses in mice.

    PubMed

    Lee, Jun Ho; Kim, Tae Hyung; Kim, Hyuk Soon; Kim, A-Ram; Kim, Do-Kyun; Nam, Seung Taek; Kim, Hyun Woo; Park, Young Hwan; Her, Erk; Park, Yeong Min; Kim, Hyung Sik; Kim, Young Mi; Choi, Wahn Soo

    2015-06-15

    Mast cells, constituents of virtually all organs and tissues, are critical cells in IgE-mediated allergic responses. The aim of this study was to investigate the effect and mechanism of an indoxyl chromogenic compound, 5-bromo-4-chloro-3-indolyl 1,3-diacetate, CAC-0982, on IgE-mediated mast cell activation and allergic responses in mice. CAC-0982 reversibly suppressed antigen-stimulated degranulation in murine mast cells (IC50, ~3.8?M) and human mast cells (IC50, ~3.0?M). CAC-0982 also inhibited the expression and secretion of IL-4 and TNF-? in mast cells. Furthermore, CAC-0982 suppressed the mast cell-mediated allergic responses in mice in a dose-dependent manner (ED50 27.9mg/kg). As for the mechanism, CAC-0982 largely suppressed the phosphorylation of Syk and its downstream signaling molecules, including LAT, Akt, Erk1/2, p38, and JNK. Notably, the tyrosine kinase assay of antigen-stimulated mast cells showed that CAC-0982 inhibited Fyn kinase, one of the upstream tyrosine kinases for Syk activation in mast cells. Taken together, these results suggest that CAC-0982 may be used as a new treatment for regulating IgE-mediated allergic diseases through the inhibition of the Fyn/Syk pathway in mast cells. PMID:25902337

  4. Theoretical study on the effects of a 4,6-O-diacetal protecting group on the stability of ion pairs from d-mannopyranosyl and d-glucopyranosyl triflates.

    PubMed

    Hosoya, Takashi; Kosma, Paul; Rosenau, Thomas

    2015-06-26

    Ion pair formation from 2,3-di-O-methyl-4,6-O-formylidene-?-d-mannopyranosyl triflate ?TMan and its d-glucopyranosyl counterpart ?TGlc was investigated at the DFT(M06-2X) level of theory, for the purpose of clarifying the effects of the 4,6-tethering on the structure and stability of ?- and ?-contact ion pairs and solvent-separated ion pairs at -78 °C. In both mannopyranosyl and glucopyranosyl triflates, the 4,6-O-formylidene group destabilized (4)H3-type conformers of the ?-contact ion pairs, rendering B2,5-types the exclusive conformers for this species. The B2,5-like ?-contact ion pair from ?TMan was 3.5 kcal/mol more stable than that from ?TGlc, probably due to the stabilizing effect by the planar O-5-C-1-C-2-O-2 dihedral angle of the former. This difference in stability of the ?-contact ion pair between the mannopyranosyl and glucopyranosyl series gives insights into the mechanisms underlying the reported experimental observation that highly ?-selective mannosylation can be achieved with 4,6-O-diacetal mannopyranosyl donors. PMID:25981459

  5. Controlled silver-staining of nucleolus organizer regions with a protective colloidal developer: a 1-step method

    Microsoft Academic Search

    W. M. Howell; D. A. Black

    1980-01-01

    Summary A 1-step silver-staining technique, requiring only 2 min to perform, is described for the differential staining of nucleolus organizer regions. A protective colloidal developer is used to control the reduction of the silver.

  6. Positive and Negative Staining of Viruses on TEM Grids Jennifer Brum, Tucson Marine Phage Lab

    E-print Network

    Sullivan, Matthew B.

    1 Positive and Negative Staining of Viruses on TEM Grids Jennifer Brum, Tucson Marine Phage Lab-cap tube - this removes any particles of uranyl acetate that have not fully dissolved 3) Fill 3x 2 ml screw

  7. Differentiation of cartilage and bone in human fetal temporal bones with Luxol fast blue stain.

    PubMed

    Declau, F; Moeneclaey, L; Forton, G; Marquet, J

    1988-01-01

    A modified Luxol fast blue technique was used to study the development of the temporal bone. This staining method makes it possible to make a clear distinction between the primitive cartilage present and the new forming bone. Although these tissues both contain a significant amount of collagen, their staining properties with the Luxol dyes are widely dissimilar, due to the different physicochemical properties of the collagen types involved in these tissues. The differentiation of mesenchymatous tissue into ligaments and joints can also be very clearly demonstrated with this technique. In studying the endochondral ossification process of the otic capsule and middle ear, the modified Luxol fast blue stain is a valuable technique that is complementary to more conventional staining methods. PMID:2460074

  8. Molecular Identification of Leishmania Species Using Samples Obtained from Negative Stained Smears

    PubMed Central

    Mohaghegh, MA; Fata, A; Salehi, GH; Berenji, F; bazzaz, M Mousavi; Rafatpanah, H; Parian, M; Movahedi, A

    2013-01-01

    Background Cutaneous Leishmaniasis (CL) is a parasitic skin disease. Diagnosis primarily is based on clinical signs and microscopic observation of parasite on direct stained smears or tissue sections. Sensitivity of direct smear is not as high as molecular methods. The aim of this study was to identify and characterize Leishmania species among the negative direct smears obtained from skin ulcers suspected to CL by PCR method. Methods Among 81 patients with suspicious skin lesions to CL referred to the Parasitology lab, negative Giemsa stained smears were collected. DNA extraction performed by scraping stained smears, then PCR was performed. Results Among the DNA extracted from smears, L. tropica was isolated from 9 (11.1%) of the smears and L.major was not isolated from any samples. Conclusion Direct microscopy on stained smears for diagnosis of leishmaniasis is not enough accurate. PCR is recommended for clinically suspected lesions with negative result of direct smear. PMID:23914250

  9. Might the Masson trichrome stain be considered a useful method for categorizing experimental tendon lesions?

    PubMed

    Martinello, Tiziana; Pascoli, Francesco; Caporale, Giovanni; Perazzi, Anna; Iacopetti, Ilaria; Patruno, Marco

    2015-08-01

    Strain injuries of tendons are the most common orthopedic injuries in athletic subjects, be they equine or human. When the tendon is suddenly damaged, an acute inflammatory phase occurs whereas its repetitive overloading may cause chronic injuries. Currently the criteria used for grading injuries are general and subjective, and therefore a reliable grading method would be an improvement. The main purpose of this study was to assess qualitatively the histological pattern of Masson trichrome stain in healthy and injured tendons; indeed, the known "paradox" of Masson staining was used to create an evaluation for the matrix of tendons, following experimental lesions and natural repair processes. A statistically significant difference of aniline-staining between healthy and lesioned tendons was observed. Overall, we think that the Masson staining might be regarded as an informative tool in discerning the collagen spatial arrangement and therefore the histological characteristics of tendons. PMID:25733060

  10. Embedding, serial sectioning and staining of zebrafish embryos using JB-4 resin.

    PubMed

    Sullivan-Brown, Jessica; Bisher, Margaret E; Burdine, Rebecca D

    2011-01-01

    Histological techniques are critical for observing tissue and cellular morphology. In this paper, we outline our protocol for embedding, serial sectioning, staining and visualizing zebrafish embryos embedded in JB-4 plastic resin-a glycol methacrylate-based medium that results in excellent preservation of tissue morphology. In addition, we describe our procedures for staining plastic sections with toluidine blue or hematoxylin and eosin, and show how to couple these stains with whole-mount RNA in situ hybridization. We also describe how to maintain and visualize immunofluorescence and EGFP signals in JB-4 resin. The protocol we outline-from embryo preparation, embedding, sectioning and staining to visualization-can be accomplished in 3 d. Overall, we reinforce that plastic embedding can provide higher resolution of cellular details and is a valuable tool for cellular and morphological studies in zebrafish. PMID:21212782

  11. Histochemical fluorescent staining of Sendai virus-infected cells with a novel sialidase substrate.

    PubMed

    Takano, Maiko; Takahashi, Tadanobu; Agarikuchi, Takashi; Kurebayashi, Yuuki; Minami, Akira; Otsubo, Tadamune; Ikeda, Kiyoshi; Kanazawa, Hiroaki; Suzuki, Takashi

    2014-09-01

    Sialidases, enzymes that remove terminal sialic acid residues, are pivotal in various biological processes such as malignancy and infection with pathogens. For histochemical staining of sialidase activity, we have developed a new synthetic sialidase substrate, sialic acid-conjugated fluorescent benzothiazolylphenol derivative (BTP3-Neu5Ac), for rapid, sensitive, and specific fluorescent staining of sialidase activity. Here, we showed the usefulness of BTP3-Neu5Ac for histochemical fluorescent staining of cells infected with Sendai virus (SV), which possesses sialidase activity. BTP3-Neu5Ac also visualised SV-infected regions of lung sections from SV-infected mice. We succeeded in histochemical fluorescent staining of SV both in vitro and in vivo. SV has been utilised in many virological and biotechnological studies such as developments of an oncolytic virus, a gene therapy vector, and a vaccine candidate. BTP3-Neu5Ac should contribute to rapid progress of such studies and researches on viral sialidase. PMID:25090482

  12. Influence of the temperature on the detection of fluorescent Y-bodies in blood stains.

    PubMed

    Thomsen, J L

    1978-01-01

    An investigation on the significance of the temperature when examining the presence of Y-bodies in cells from blood stains was performed. Stains on cotton cloth were placed at 53 degrees C and 5 degrees C respectively, and the results were compared with those of an earlier report on stains stored at room temperature. There proved to be a much more rapid decline of the male count, and false negative results appeared earlier. This was most pronounced for the "cold" stains. No false positives were found. Twenty-five fresh blood smears from one male were examined in order to get an impression of the accidental variation. It was found that even in the case of a male with a rather low count (mean: 28%) the chance of accidentally getting a false negative result (less than 10%) was less than 2%. PMID:658857

  13. Assessing Nezara viridula (Hemiptera: Pentatomidae) feeding damage in macadamia nuts by using a biological stain.

    PubMed

    Golden, Mary; Follett, Peter A; Wright, Mark G

    2006-06-01

    Damage caused by southern green stink bug, Nezara viridula (L.), to macadamia nuts, Macadamia integrifolia Maiden & Betche, is normally determined after nuts are harvested and processed, which may be many months after damage occurred in the field. We developed a method using ruthenium red dye to stain stink bug feeding probes and indirectly assess feeding activity in macadamia nuts. By using the staining method, feeding probes were easily detected on the husk, shell, and kernel. Husk probing was highly correlated (0.80-0.90) with feeding and damage to the kernel. Failure rate to detect kernel damage from stained husk probes was generally <6%. The staining method was equally effective for immature and mature nuts; therefore, N. viridula feeding activity can be monitored throughout the season to evaluate pest management tactics and forecast outbreak populations. PMID:16813317

  14. Limonite-stained Siltite near the Blackbird Cobalt-Copper Mine

    USGS Multimedia Gallery

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

  15. Transient partial permeabilization with saponin enables cellular barcoding prior to surface marker staining1

    PubMed Central

    Behbehani, Gregory K.; Thom, Colin; Zunder, Eli R.; Finck, Rachel; Gaudilliere, Brice; Fragiadakis, Gabriela K.; Fantl, Wendy J.; Nolan, Garry P.

    2015-01-01

    Fluorescent cellular barcoding and mass-tag cellular barcoding are cytometric methods that enable high sample throughput, minimize inter-sample variation, and reduce reagent consumption. Previously employed barcoding protocols require that barcoding be performed after surface marker staining, complicating combining the technique with measurement of alcohol-sensitive surface epitopes. This report describes a method of barcoding fixed cells after a transient partial permeabilization with 0.02% saponin that results in efficient and consistent barcode staining with fluorescent or mass-tagged reagents while preserving surface marker staining. This approach simplifies barcoding protocols and allows direct comparison of surface marker staining of multiple samples without concern for variations in the antibody cocktail volume, antigen-antibody ratio, or machine sensitivity. Using this protocol, cellular barcoding can be used to reliably detect subtle differences in surface marker expression. PMID:25274027

  16. Staining Pattern Classification of Antinuclear Autoantibodies Based on Block Segmentation in Indirect Immunofluorescence Images

    PubMed Central

    Li, Jiaqian; Tseng, Kuo-Kun; Hsieh, Zu Yi; Yang, Ching Wen; Huang, Huang-Nan

    2014-01-01

    Indirect immunofluorescence based on HEp-2 cell substrate is the most commonly used staining method for antinuclear autoantibodies associated with different types of autoimmune pathologies. The aim of this paper is to design an automatic system to identify the staining patterns based on block segmentation compared to the cell segmentation most used in previous research. Various feature descriptors and classifiers are tested and compared in the classification of the staining pattern of blocks and it is found that the technique of the combination of the local binary pattern and the k-nearest neighbor algorithm achieve the best performance. Relying on the results of block pattern classification, experiments on the whole images show that classifier fusion rules are able to identify the staining patterns of the whole well (specimen image) with a total accuracy of about 94.62%. PMID:25474260

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

    ERIC Educational Resources Information Center

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

    2002-01-01

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

  18. High contrast en bloc staining of neuronal tissue for field emission scanning electron microscopy

    PubMed Central

    Tapia, Juan C.; Kasthuri, Narayanan; Hayworth, Kenneth; Schalek, Richard; Lichtman, Jeff W.; Smith, Stephen J; Buchanan, JoAnn

    2013-01-01

    Conventional heavy metal post staining methods on thin sections lend contrast but often cause contamination. To avoid this problem, we tested several en bloc staining techniques to contrast tissue in serial sections mounted on solid substrates for examination by Field Emission Scanning Electron Microscope (FESEM). Because FESEM section imaging requires that specimens have higher contrast and greater electrical conductivity than transmission electron microscope (TEM) samples, our technique utilizes osmium impregnation (OTO) to make the samples conductive while heavily staining membranes for segmentation studies. Combining this step with other classic heavy metal en bloc stains including uranyl acetate, lead aspartate, copper sulfate and lead citrate produced clean, highly contrasted TEM and SEM samples of insect, fish, and mammalian nervous system. This protocol takes 7–15 days to prepare resin embedded tissue, cut sections and produce serial section images. PMID:22240582

  19. Cutaneous New World Leishmaniasis on a Port-wine stain birthmark*

    PubMed Central

    Criado, Paulo Ricardo; Valente, Neusa Sakai; Noda, Aliene; Belda, Walter

    2014-01-01

    We present an interesting case report of two sarcoid-like lesions on a port-wine stain (PWS) birthmark in a Brazilian patient which on investigation proved to be cutaneous leishmaniasis. PMID:25054762

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

    E-print Network

    Darrouzet, Stephen 1987-

    2011-04-21

    analysis of the vessel’s response to the implant with greater sensitivity and specificity while reducing beam-hardening artifact from stent struts. The developed staining techniques included iodine-potassium iodide, phosphomolybdic acid...

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

    SciTech Connect

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

    1992-01-01

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

  2. Simplified heavy metal staining techniques demonstrated with Fast Plant leaf tissue

    Microsoft Academic Search

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

    1992-01-01

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

  3. Silver-staining of proteins in polyacrylamide gels: a general overview

    Microsoft Academic Search

    Thierry Rabilloud; L. Vuillard; C. Gilly; J. J. Lawrence

    2009-01-01

    On the basis of the physico-chemical principles underlying silver-staining of proteins, which are recalled in this paper, several methods of silver-staining of proteins after SDS electrophoresis in polyacrylamide gels or isoelectric focusing were tested. The most valuable protocols are presented in this report, including standard methods for unsupported gels and new methods devised for thin (0.5 mm) supported gels for

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

    Microsoft Academic Search

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

    1961-01-01

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

  5. Staining of proteins on SDS polyacrylamide gels and on nitrocellulose membranes by Alta, a colour used as a cosmetic

    Microsoft Academic Search

    Jayanta K. Pal; Dhanashri Godbole; Kiran Sharma

    2004-01-01

    We describe here the use of Alta, a pre-existing scarlet-red stain of cosmetic use, for staining proteins on sodium dodecyl sulfate (SDS) polyacrylamide gels, as well as for a single step staining of gels and nitrocellulose membranes during Western blot analysis. This stain, which is composed of 0.8% Crocein scarlet (brilliant crocein) and 0.2% Rhodamine B, is inexpensive, easy to

  6. [Evaluation of visualization of biological stains with the use of alternative light source (ALS) for the purpose of genetic identification. Part I. Blood and saliva stains analysis].

    PubMed

    Szeremeta, Micha?; Pepi?ski, Witold; Niemcunowicz-Janica, Anna; Skawro?ska, Ma?gorzata; Sackiewicz, Adam; Ptaszy?ska-Sarosiek, Iwona; Ok?ota, Magdalena

    2010-01-01

    The objective of the investigation was evaluation of visualization of human blood and saliva stains with the use of alternative light source for the purpose of genetic identification. Experimental bloodstains on the bright base were the most clearly seen in the natural light and white light, up to blood dilution of 1:600. Complete typeability of AmpFISTR SGM Plus kit profiles was obtained from bloodstains at dilution 1:1500. Partial AmpFISTR SGM Plus kit profiles were typed from bloodstains at dilutions 1:1750 and 1:2000. Experimental saliva stains on the light-colored base were completely invisible in the natural light and white light, while they were visualized at wavelength range 300-415 nm through yellow goggles, and at wavelength range 300-455 nm through orange goggles at saliva dilution 1: 600. Complete typeability of AmpFISTR SGM Plus kit loci was obtained from saliva stains at dilution 1:1750. Partial AmpFISTR SGM Plus kit profiles were typed from saliva stains at dilution 1:2000. The wavelength of 455 nm and orange goggles were the optimal set for visualization of bloodstains on various, noncontrasting materials. Other useful wavelength/combinations of goggles were CSS light/red goggles. In case of saliva, the most useful general condition for visualization of stains on various, non-contrasting materials was with the wavelength set to 300-415 nm, while wearing yellow goggles. Other useful combinations of wavelength/goggles were 300-455 nm/orange or red goggles, and also CSS light/orange or red goggles. PMID:21863732

  7. Cell wall staining with Trypan blue enables quantitative analysis of morphological changes in yeast cells

    PubMed Central

    Liesche, Johannes; Marek, Magdalena; Günther-Pomorski, Thomas

    2015-01-01

    Yeast cells are protected by a cell wall that plays an important role in the exchange of substances with the environment. The cell wall structure is dynamic and can adapt to different physiological states or environmental conditions. For the investigation of morphological changes, selective staining with fluorescent dyes is a valuable tool. Furthermore, cell wall staining is used to facilitate sub-cellular localization experiments with fluorescently-labeled proteins and the detection of yeast cells in non-fungal host tissues. Here, we report staining of Saccharomyces cerevisiae cell wall with Trypan Blue, which emits strong red fluorescence upon binding to chitin and yeast glucan; thereby, it facilitates cell wall analysis by confocal and super-resolution microscopy. The staining pattern of Trypan Blue was similar to that of the widely used UV-excitable, blue fluorescent cell wall stain Calcofluor White. Trypan Blue staining facilitated quantification of cell size and cell wall volume when utilizing the optical sectioning capacity of a confocal microscope. This enabled the quantification of morphological changes during growth under anaerobic conditions and in the presence of chemicals, demonstrating the potential of this approach for morphological investigations or screening assays. PMID:25717323

  8. Issues in using whole slide imaging for diagnostic pathology: "routine" stains, immunohistochemistry and predictive markers.

    PubMed

    Taylor, C R

    2014-08-01

    The traditional microscope, together with the "routine" hematoxylin and eosin (H & E) stain, remains the "gold standard" for diagnosis of cancer and other diseases; remarkably, it and the majority of associated biological stains are more than 150 years old. Immunohistochemistry has added to the repertoire of "stains" available. Because of the need for specific identification and even measurement of "biomarkers," immunohistochemistry has increased the demand for consistency of performance and interpretation of staining results. Rapid advances in the capabilities of digital imaging hardware and software now offer a realistic route to improved reproducibility, accuracy and quantification by utilizing whole slide digital images for diagnosis, education and research. There also are potential efficiencies in work flow and the promise of powerful new analytical methods; however, there also are challenges with respect to validation of the quality and fidelity of digital images, including the standard H & E stain, so that diagnostic performance by pathologists is not compromised when they rely on whole slide images instead of traditional stained tissues on glass slides. PMID:24325681

  9. A six-week clinical study to compare the stain removal efficacy of three dentifrices.

    PubMed

    Nathoo, Salim; Petrone, Margaret E; DeVizio, William; Chaknis, Patricia; Volpe, Anthony R

    2002-01-01

    The objective of this double-blind clinical study was to compare the tooth whitening efficacy (stain removal) of a new commercially available tooth whitening dentifrice (Colgate Total Plus Whitening Toothpaste) containing 0.2% triclosan and 3.0% PVM/MA copolymer in a 0.243% sodium fluoride/high cleaning silica base, with that of two commercially available dentifrices, Crest Multi-Care Advanced Cleaning Toothpaste and Colgate Winterfresh Gel Fluoride Toothpaste. Following a baseline examination to assess extrinsic tooth stain, qualifying adult male and female subjects were randomized into three treatment groups which were balanced for gender, age and level of extrinsic tooth stain. Subjects were asked to brush their teeth twice (morning and evening) for one minute with their assigned dentifrice using a soft-bristled toothbrush. Examinations for extrinsic tooth stain were repeated after six weeks' use of the study dentifrices. One-hundred and twenty-three (123) subjects complied with the protocol and completed the study. At the six-week examination, subjects assigned to the Colgate Total Plus Whitening Toothpaste treatment group exhibited statistically significant reductions in extrinsic tooth stain area and extrinsic tooth stain intensity relative to those subjects assigned to the Crest Multi-Care Advanced Cleaning Toothpaste and the Colgate Winterfresh Gel Fluoride Toothpaste. PMID:11695214

  10. Rapid, one step staining procedures for analysis of cellular DNA and protein by single and dual laser flow cytometry

    Microsoft Academic Search

    H. A. Crissman; J. A. Steinkamp

    1982-01-01

    Detailed, simplified techniques are described for simultaneous staining and analysis of DNA and protein in a number of mammalian cell types. Cell staining involves the addition of appropriate dye mixtures to unfixed or ethanol-fixed cells and subsequent analysis of cell populations in the staining reagents generally within 10 to 20 minutes. The approach is novel in that no centrifugation steps

  11. Lasers in Surgery and Medicine 39:118127 (2007) Laser Surgery of Port Wine Stains Using Local Vaccum

    E-print Network

    Aguilar, Guillermo

    2007-01-01

    Lasers in Surgery and Medicine 39:118­127 (2007) Laser Surgery of Port Wine Stains Using Local port wine stain (PWS) laser treatment. Study Design/ Materials and Methods: Mathematical models INTRODUCTION The goal of cutaneous laser surgery during treatment of port wine stain (PWS) birthmarks

  12. Experimental study of multiple-intermittent cryogen spurts and laser pulses for the treatment of port wine stain birthmarks

    E-print Network

    Aguilar, Guillermo

    of port wine stain birthmarks Guillermo Aguilar1,2 , Bernard Choi1 , John A. Viator1 , Dan Andersen3 of epidermal damage during pulsed laser treatment of port wine stain (PWS) birthmarks. Unfortunately such as port wine stain (PWS) birthmarks (150 to 500 µm deep) [1-2]. Clinical studies [2-4] have demonstrated

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

    E-print Network

    Paris-Sud XI, Université de

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

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

    PubMed

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

    2014-12-01

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

  15. Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels

    PubMed Central

    Dyballa, Nadine; Metzger, Sabine

    2009-01-01

    Coomassie Brilliant Blue (CBB) is a dye commonly used for the visualization of proteins separated by SDS-PAGE, offering a simple staining procedure and high quantitation. Furthermore, it is completely compatible with mass spectrometric protein identification. But despite these advantages, CBB is regarded to be less sensitive than silver or fluorescence stainings and therefore rarely used for the detection of proteins in analytical gel-based proteomic approaches. Several improvements of the original Coomassie protocol1 have been made to increase the sensitivity of CBB. Two major modifications were introduced to enhance the detection of low-abundant proteins by converting the dye molecules into colloidal particles: In 1988, Neuhoff and colleagues applied 20% methanol and higher concentrations of ammonium sulfate into the CBB G-250 based staining solution2, and in 2004 Candiano et al. established Blue Silver using CBB G-250 with phosphoric acid in the presence of ammonium sulfate and methanol3. Nevertheless, all these modifications just allow a detection of approximately 10 ng protein. A widely fameless protocol for colloidal Coomassie staining was published by Kang et al. in 2002 where they modified Neuhoff's colloidal CBB staining protocol regarding the complexing substances. Instead of ammonium sulfate they used aluminum sulfate and methanol was replaced by the less toxic ethanol4. The novel aluminum-based staining in Kang's study showed superior sensitivity that detects as low as 1 ng/band (phosphorylase b) with little sensitivity variation depending on proteins. Here, we demonstrate application of Kang's protocol for fast and sensitive colloidal Coomassie staining of proteins in analytical purposes. We will illustrate the quick and easy protocol using two-dimensional gels routinely performed in our working group. PMID:19684561

  16. Fast and sensitive colloidal coomassie G-250 staining for proteins in polyacrylamide gels.

    PubMed

    Dyballa, Nadine; Metzger, Sabine

    2009-01-01

    Coomassie Brilliant Blue (CBB) is a dye commonly used for the visualization of proteins separated by SDS-PAGE, offering a simple staining procedure and high quantitation. Furthermore, it is completely compatible with mass spectrometric protein identification. But despite these advantages, CBB is regarded to be less sensitive than silver or fluorescence stainings and therefore rarely used for the detection of proteins in analytical gel-based proteomic approaches. Several improvements of the original Coomassie protocol(1) have been made to increase the sensitivity of CBB. Two major modifications were introduced to enhance the detection of low-abundant proteins by converting the dye molecules into colloidal particles: In 1988, Neuhoff and colleagues applied 20% methanol and higher concentrations of ammonium sulfate into the CBB G-250 based staining solution(2), and in 2004 Candiano et al. established Blue Silver using CBB G-250 with phosphoric acid in the presence of ammonium sulfate and methanol(3). Nevertheless, all these modifications just allow a detection of approximately 10 ng protein. A widely fameless protocol for colloidal Coomassie staining was published by Kang et al. in 2002 where they modified Neuhoff's colloidal CBB staining protocol regarding the complexing substances. Instead of ammonium sulfate they used aluminum sulfate and methanol was replaced by the less toxic ethanol(4). The novel aluminum-based staining in Kang's study showed superior sensitivity that detects as low as 1 ng/band (phosphorylase b) with little sensitivity variation depending on proteins. Here, we demonstrate application of Kang's protocol for fast and sensitive colloidal Coomassie staining of proteins in analytical purposes. We will illustrate the quick and easy protocol using two-dimensional gels routinely performed in our working group. PMID:19684561

  17. Al adjuvants can be tracked in viable cells by lumogallion staining.

    PubMed

    Mile, Irene; Svensson, Andreas; Darabi, Anna; Mold, Matthew; Siesjö, Peter; Eriksson, Håkan

    2015-07-01

    The mechanism behind the adjuvant effect of aluminum salts is poorly understood notwithstanding that aluminum salts have been used for decades in clinical vaccines. In an aqueous environment and at a nearly neutral pH, the aluminum salts form particulate aggregates, and one plausible explanation of the lack of information regarding the mechanisms could be the absence of an efficient method of tracking phagocytosed aluminum adjuvants and thereby the intracellular location of the adjuvant. In this paper, we want to report upon the use of lumogallion staining enabling the detection of phagocytosed aluminum adjuvants inside viable cells. Including micromolar concentrations of lumogallion in the culture medium resulted in a strong fluorescence signal from cells that had phagocytosed the aluminum adjuvant. The fluorescence appeared as spots in the cytoplasm and by confocal microscopy and co-staining with probes presenting fluorescence in the far-red region of the spectrum, aluminum adjuvants could to a certain extent be identified as localized in acidic vesicles, i.e., lysosomes. Staining and detection of intracellular aluminum adjuvants was achieved not only by diffusion of lumogallion into the cytoplasm, thereby highlighting the presence of the adjuvant, but also by pre-staining the aluminum adjuvant prior to incubation with cells. Pre-staining of aluminum adjuvants resulted in bright fluorescent particulate aggregates that remained fluorescent for weeks and with only a minor reduction of fluorescence upon extensive washing or incubation with cells. Both aluminum oxyhydroxide and aluminum hydroxyphosphate, two of the most commonly used aluminum adjuvants in clinical vaccines, could be pre-stained with lumogallion and were easily tracked intracellularly after incubation with phagocytosing cells. Staining of viable cells using lumogallion will be a useful method in investigations of the mechanisms behind aluminum adjuvants' differentiation of antigen-presenting cells into inflammatory cells. Information will be gained regarding the phagosomal pathways and the events inside the phagosomes, and thereby the ultimate fate of phagocytosed aluminum adjuvants could be resolved. PMID:25896212

  18. The usefulness of changing focus during examination using Gram staining as initial diagnostic clue for infective tuberculosis.

    PubMed

    Atsukawa, Yoshiko; Kawakami, Sayoko; Asahara, Miwa; Ishigaki, Shinobu; Tanaka, Takashi; Ono, Yasuo; Nishiya, Hajime; Fujisaki, Ryuichi; Koga, Ichiro; Ota, Yasuo; Miyazawa, Yukihisa

    2011-08-01

    Gram staining is a useful technique for detecting bacteria but is highly questionable in detecting Mycobacterium tuberculosis. Its detection generally requires special staining, such as Ziehl-Neelsen staining. We experienced three cases in which tuberculosis was first suggested by Gram staining of sputum or pus, confirmed by Ziehl-Neelsen staining, and diagnosed by polymerase chain reaction or culture. To find colorless tubercle bacilli in clinical samples with various organisms, varying the focus to slightly longer and shorter during study of the slides is indispensable. We present criteria for detecting infective pulmonary tuberculosis in Gram staining. First, in the ordinary focus, weakly stained, thin, gram-positive bacilli are found; second, with a slightly longer focus distance, the thin, cord-like, conspicuous gram-positive bacilli can be observed; and third, with a shorter focus distance, the gram-positive bacilli have changed into the brightened, colorless, or ghost ones. Four laboratory technologists each evaluated 20 Gram-stained samples after being lectured on the criteria, with no prior information about the sample. They accurately evaluated the presence of the bacilli in Gram-stained preparations in more than 90% of samples containing 3+ bacilli on Ziehl-Neelsen staining. Gram staining is available as an easy and rapid initial clue to recognize highly infective tuberculosis. PMID:21327691

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

    PubMed Central

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

    2012-01-01

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

  20. Characterisation of foxing stains in eighteenth to nineteenth century drawings using non-destructive techniques.

    PubMed

    Manso, M; Pessanha, S; Figueira, F; Valadas, S; Guilherme, A; Afonso, M; Rocha, A C; Oliveira, M J; Ribeiro, I; Carvalho, M L

    2009-12-01

    The reddish-brown, brown or yellowish stains of circular or irregular shape known as foxing spots have been fully described in conservation literature but still, this phenomenon does not find any scientific agreement since many hypotheses have been raised concerning their origin. In this work a contribution to foxing definition not only focussed on its appearance but also reported on its chemical information. For this purpose foxing stains present in drawings from two Portuguese artists dated from the eighteenth to nineteenth centuries were observed under ultra-violet light and optical microscope and analysed by three non-invasive spectroscopy techniques. The observations carried out on the stains provided information on their surface morphology. The use of energy-dispersive X-ray fluorescence revealed a variation on the elemental content between foxing and paper region. Although the results from X-ray diffraction analysis showed no signs of cellulose degradation in foxing stains, Fourier-transformed infrared analysis revealed the presence of oxide groups. Both the information on the chemical nature and surface morphology of the stains achieved in this study will contribute to increase foxing formation information and develop future protocols for conservation purposes. PMID:19784831

  1. The choice of a masking agent in the histochemical staining of metals.

    PubMed

    Sumi, Y; Muraki, T; Suzuki, T

    1983-03-01

    The masking effects of standard masking agents (aminopolycarboxylic acids, carboxylic acids and phosphates) have been investigated in both test-tube experiments and tissue sections in order to ascertain the factors which must be considered when choosing a masking agent for the histochemical staining of a metal. The masking effects in vitro were determined by spectrophotometry through the complexing of the dye Chrome Azurol S with aluminium, beryllium, and iron at pH 5 and 7. The effects were also examined by staining metal-containing tissue sections in a Chrome Azurol S masking agent system at the same pH values. In many cases, the masking effects observed in sections did not agree with those obtained in the test-tube experiments. This means that the published values of stability constants are not a sufficient guide for choosing a suitable masking agent for the staining of metals. The discrepancy is mainly attributable to the presence of protein in a solid state when metals are stained in sections. Therefore, in the future, consideration should be given to a metal-protein or masking agent-protein interaction using a model compound such as a chelate resin. The polyphosphates are among the most useful masking agents for metal staining in acidic solutions from a practical standpoint. PMID:6189808

  2. R-phycoerythrin-conjugated antibodies are inappropriate for intracellular staining of murine plasma cells.

    PubMed

    Kim, Myun Soo; Kim, Tae Sung

    2013-05-01

    Phycoerythrin (PE) is a type of phycobiliproteins found in cyanobacteria and red algae. PE-conjugated antibodies are broadly used for flow cytometry and immunofluorescence microscopy. Because nonspecific binding of antibodies results in decreased analytic accuracy, numerous efforts have been made to unveil cases and mechanisms of nonspecific bindings. However, nonspecific binding of specific cell types by a fluorescent dye-conjugated form of antibody has been rarely reported. In the present study, we discovered that PE-conjugated antibodies, but not FITC- or APC-antibodies, selectively stained lamina propria plasma cells (LP-PCs) from the murine small intestine after membrane permeabilization. We demonstrated that LP-PC-selective staining with PE-antibodies was not due to interactions of antibody-epitope or antibody-Fc receptor. This unexpected staining by PE-antibody was not dependent on the mouse strain of LP-PCs, experimental methods, or origin species of the antibody, but dependent on PE itself. This phenomenon was also observed in plasma cells isolated from bone marrow, spleen, and mesenteric lymph nodes. Furthermore, in vitro activated B cells and in vivo generated LP-PCs were also selectively stained by PE-conjugated antibodies. Taken together, these results show that PE-conjugated antibodies are inappropriate for intracellular staining of murine plasma cells. PMID:23463627

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

    SciTech Connect

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

    1985-01-01

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

  4. Staining intensity of individual osteons correlated with elastic properties and degrees of mineralization.

    PubMed

    Qin, L; Hung, L; Leung, K; Guo, X; Bumrerraj, S; Katz, L

    2001-01-01

    Surface staining is widely used for histological studies involving undecalcified thick bone sections. Individual osteons, particularly newly formed ones stained with toluidine blue (TB), show various color intensities. We studied the correlations between TB color intensity and the differences in stiffness and degree of mineralization of individual osteons in undecalcified histological sections of goat tibial diaphysis, measured by scanning acoustic microscopy (SAM) and contact microradiography (CMR), respectively. Results showed that all three measurements correlated significantly with each other (r = 0.567 - 0.786; all P < or = 0.01). The TB surface staining intensity of individual osteons correlated better with the reflection coefficient (stiffness index) measured by SAM (r = 0.713) than with the aluminum step-wedge equivalent thickness measured on CMR micrographs (r = 0.567). The aluminum step-wedge equivalent thickness of individual osteons on CMR correlated slightly better with the SAM reflection coefficient (r = 0.786) than with the TB surface staining intensity (r = 0.713). The results of this study suggest that TB surface staining may be used as a simple method for indicating differences in stiffness and degree of mineralization in individual osteons in comparative histological studies. PMID:11685651

  5. Effect of hydrogen peroxide mouthwash as an adjunct to chlorhexidine on stains and plaque

    PubMed Central

    Jhingta, Pravesh; Bhardwaj, Ashu; Sharma, Deepak; Kumar, Naresh; Bhardwaj, Vinay Kumar; Vaid, Sanjeev

    2013-01-01

    Aim: To investigate whether the use of an oxidizing mouth rinse as an adjunct to chlorhexidine is efficacious in reducing stains and plaque. Materials and Methods: This study had a single-blind, three-group (n = 35 each) parallel design, including a 21 days experimental period during which group I rinsed with chlorhexidine (CHX) 0.2% alone, group II used chlorhexidine (CHX) followed by hydrogen peroxide (H2O2) 1.5%. Group III rinsed with the same mouthwashes in reverse order. Patients were randomly assigned to one of the three groups. The examination for plaque, and stains was done after 1, 2, and 3 weeks of rinsing. Results: Group II showed significantly less stain intensity in comparison with group I after 14 and 21 days (P values 0.025 and 0.005, respectively). The proportion of stained surfaces was less in the group II than in the group I and was significant at the end of 1 week. The plaque formation was significantly less in groups II and III than group I at 7, 14, and 21 days. Conclusion: The adjunctive use of hydrogen peroxide to chlorhexidine proved to be superior to chlorhexidine alone with regard to the inhibition of plaque and development of stains. PMID:24174723

  6. Diagnostic value of immunohistochemical staining of GP73, GPC3, DCP, CD34, CD31, and reticulin staining in hepatocellular carcinoma.

    PubMed

    Yao, Shuzhe; Zhang, Jianping; Chen, Haiyan; Sheng, Yan; Zhang, Xiaoying; Liu, Zhiyan; Zhang, Cuijuan

    2013-09-01

    It has been reported that Golgi protein-73 (GP73), glypican-3 (GPC3), and des-?-carboxy prothrombin (DCP) could serve as serum markers for the early detection of hepatocellular carcinoma (HCC). This study aimed to evaluate a panel of immunostaining markers (including GP73, GPC3, DCP, CD34, and CD31) as well as reticulin staining to distinguish HCC from the mimickers. Our results revealed that CD34 immunostaining and reticulin staining were highly sensitive for the diagnosis of HCC. A special immunoreaction pattern of GP73--a diffuse coarse-block pattern in a perinuclear region or a concentrated cluster-like or cord-like pattern in a certain part of the cytoplasm--was observed in HCC cells, in contrast to the cytoplasmic fine-granular pattern in surrounding non-tumor cells and non-malignant nodules. This coarse-block pattern correlated significantly with less differentiated HCC. In comparison, GPC3 displayed a good advantage in diagnosing well-differentiated HCC. In our study, DCP and CD31 showed little diagnostic value for HCC as an immunostaining marker. When GP73, GPC3, and CD34 were combined, the specificity improved to 96.6%. Our findings demonstrate for the first time that the immunohistochemical panel of GP73, GPC3, and CD34 as well as reticulin staining is highly specific for the pathological diagnosis of HCC. PMID:23686365

  7. Morphological significance of cladosporium contaminants on materials and utensils in contact with food.

    PubMed

    Ohta, Toshiko; Park, Bong Joo; Aihara, Maki; Ri, Noritoshi; Saito, Toshiko; Sawada, Takuo; Takatori, Kosuke

    2006-06-01

    Cladosporium contaminants on materials and utensils that come into contact with food were morphologically investigated. The most common contaminants, C. cladosporioides and C. sphaerospermum, were detected on the samples. The morphological changes of the Cladosporium species were investigated by using stereoscopic, optical light, fluorescent, and scanning electron microscopes. Microscopically the Cladosporium contaminants were observed as aggregated dark brown spots, strongly pigmented, irregularly swollen, and in long chains. Using fluorescent microscopy, the Cladosporium mycelia were clearly stained with fluorescein diacetate as viable cells, but the old cells were mostly non-viable, as shown by staining with propidium iodide. The dynamics of the morphological changes showed that the penetrating mycelia were closely attached to the surface of the materials and utensils under investigation. These results provide information about the significance of Cladosporium contamination on materials and utensils in contact with food and may contribute to the control of fungal contamination. PMID:16789547

  8. New staining methods for yeast like fungi under special consideration of human pathogenic fungi

    NASA Astrophysics Data System (ADS)

    Paulitsch-Fuchs, Astrid; Treiber, Fritz; Grasser, Erik; Buzina, Walter; Rosker, Christian

    2010-11-01

    A new method for in-cellular staining of yeast like fungi with Oregon Green and SYTOX Green is presented enabling their detection as well as the observation of cellular details via confocal laser scanning microscopy. Fluorochromes play an important role in many scientific disciplines including medicine, cell biology and botany. For the visualisation of fungal cell walls Calcofluor White is the flourochrome of choice. The necessity of an UV laser for its excitation makes it unpracticable for daily use. Safranin O, DAPI, 2NBDG, Ethidium Bromide and Acridin-orange are commonly used stains for nuclei in fugal microscopy. The attention was given to the possibility of using the differences in staining patterns to distinguish certain pathogenic yeast species e.g. Candida albicans and Candida krusei. Our results show that high quality microscopy of yeast like organisms can readily be achieved by the use of two suitable fluorochromes.

  9. Identification and quantification of cooling water biofilms using fluorescent staining and ATP monitoring techniques

    SciTech Connect

    Chalut, J.; Cairns, J.; Korkorian, N. [Grace Dearborn Inc., Mississauga, Ontario (Canada)

    1994-12-31

    Biofilm formation can create corrosion problems in industrial water systems. Control of biofilms is achieved most effectively when the mechanism of formation is understood. The use of traditional microbiological analyses such as plate counts and dipslides to analyze deposits provides insufficient information about viable cell content and their role in deposit formation. The ATP assay, a newer technology, is more useful but only measures total living biomass. In order to assess the potential for biofouling in cooling water systems, novel staining and monitoring techniques have been developed. Staining technology allows characterization and assessment of biofilm composition. This staining methodology is used to complement ATP analysis of field samples. Case histories are used to illustrate the benefits of this approach. Case histories included a textile manufacturing plant, an oil refinery, and a pulp and paper mill.

  10. Stain-etched porous silicon nanostructures for multicrystalline silicon-based solar cells

    NASA Astrophysics Data System (ADS)

    Ben Rabha, M.; Hajji, M.; Belhadj Mohamed, S.; Hajjaji, A.; Gaidi, M.; Ezzaouia, H.; Bessais, B.

    2012-02-01

    In this paper, we study the optical, optoelectronic and photoluminescence properties of stain-etched porous silicon nanostructures obtained with different etching times. Special attention is given to the use of the stain-etched PS as an antireflection coating as well as for surface passivating capabilities. The surface morphology has been analyzed by scanning electron microscopy. The evolution of the Si-O and Si-H absorption bands was analyzed by Fourier transform infrared spectrometry before and after PS treatment. Results show that stain etching of the silicon surface drops the total reflectivity to about 7% in the 400-1100 nm wavelength range and the minority carrier lifetime enhances to about 48 ?s.

  11. Application of the DNA-Specific Stain Methyl Green in the Fluorescent Labeling of Embryos.

    PubMed

    Prieto, Daniel; Aparicio, Gonzalo; Machado, Matías; Zolessi, Flavio R

    2015-01-01

    Methyl green has long been known as a histological stain with a specific affinity for DNA, although its fluorescent properties have remained unexplored until recently. In this article, we illustrate the method for preparing a methyl green aqueous stock solution, that when diluted can be used as a very convenient fluorescent nuclear label for fixed cells and tissues. Easy procedures to label whole zebrafish and chick embryos are detailed, and examples of images obtained shown. Methyl green is maximally excited by red light, at 633 nm, and emits with a relatively sharp spectrum that peaks at 677 nm. It is very inexpensive, non-toxic, highly stable in solution and very resistant to photobleaching when bound to DNA. Its red emission allows for unaltered high resolution scanning confocal imaging of nuclei in thick specimens. Finally, this methyl green staining protocol is compatible with other cell staining procedures, such as antibody labeling, or actin filaments labeling with fluorophore-conjugated phalloidin. PMID:25993383

  12. Experience with the sodium sulphate-Alcian Blue stain for amyloid in cardiac pathology.

    PubMed Central

    Pomerance, A; Slavin, G; McWatt, J

    1976-01-01

    The sodium sulphate-Alcian Blue (SAB) method, which stains amyloid green, was evaluated in 220 hearts from elderly patients. The technique proved superior to the Congo red, crystal violet, and thioflavine T methods used either singly or as a battery for the demonstration of cardiac amyloid. Amyloid was easily identified under the X3 objective, even in small amounts. A few non-amyloid components stained varying shades of green but were easily distinguished on morphological grounds. No false positive or equivocal reactions occurred, and in particular elastic laminae and paravascular connective tissue were not tinctorially confused with amyloid. The SAB stain is technically simple and consistently reproducible, and no special light source is required for examination. An additional advantage in cardiac pathology is the simultaneous demonstration of any fibrosis, basophilic myofibre degeneration, tissue mast cells and mucoid degeneration of valves present. Images PMID:55419

  13. Evaluation of the staining potential of a caries infiltrant in comparison to other products.

    PubMed

    Rey, Nicolas; Benbachir, Nacer; Bortolotto, Tissiana; Krejci, Ivo

    2014-01-01

    In this study, we evaluated in vitro the staining susceptibility of an infiltration resin (Icon, DMG, Hamburg, Germany) and compared it with several marketed bonding systems. Fifty 1-mm-thick disk-shaped specimens were prepared for Icon and for each bonding material. Initial specimen color was assessed by a spectrophotometer. Specimens in each group were then randomly divided into five sub-groups and stored in an incubator at 37?C in the dark for 60 days. Groups 4 and 5 were used as negative controls by being stored dry and in tap water respectively. Test groups were stored in (1) coffee, (2) tea, or (3) red wine. After 60 days of storage, new spectrophotometric measurements were performed and dE (color difference) was calculated to determine color change. Icon showed higher staining susceptibility. The clinician should be aware of the staining potential of infiltration resins over time. PMID:24492117

  14. Vascular Engorgement of Lacrimal Gland Associated With Port-Wine Stain.

    PubMed

    Talcott, Katherine E; Lee, Nahyoung Grace; Freitag, Suzanne K

    2014-09-01

    Port-wine stains are congenital dermal capillary malformations that typically involve the head and neck. While most of them are isolated malformations, they have been associated with other vascular findings, including conjunctival, episcleral, and choroidal hemangiomas. They have also been associated with the phakomatosis Sturge-Weber syndrome, characterized by parieto-occipital, leptomeningeal, and ocular choroidal vascular malformations. However, vascular engorgement of the lacrimal gland has not been previously reported in association with port-wine stains. The authors present a case of a 52-year-old man with a long-standing and isolated right periorbital port-wine stain referred for lacrimal gland enlargement on CT scan. He was found to have asymptomatic right lacrimal gland vascular engorgement, which was radiographically stable over a period of 5 years. PMID:25198395

  15. Unsupervised content classification based nonrigid registration of differently stained histology images.

    PubMed

    Song, Y; Treanor, D; Bulpitt, A J; Wijayathunga, N; Roberts, N; Wilcox, R; Magee, D R

    2014-01-01

    Registration of histopathology images of consecutive tissue sections stained with different histochemical or immunohistochemical stains is an important step in a number of application areas, such as the investigation of the pathology of a disease, validation of MRI sequences against tissue images, multiscale physical modeling, etc. In each case, information from each stain needs to be spatially aligned and combined to ascertain physical or functional properties of the tissue. However, in addition to the gigabyte-size images and nonrigid distortions present in the tissue, a major challenge for registering differently stained histology image pairs is the dissimilar structural appearance due to different stains highlighting different substances in tissues. In this paper, we address this challenge by developing an unsupervised content classification method that generates multichannel probability images from a roughly aligned image pair. Each channel corresponds to one automatically identified content class. The probability images enhance the structural similarity between image pairs. By integrating the classification method into a multiresolution-block-matching-based nonrigid registration scheme (N. Roberts, D. Magee, Y. Song, K. Brabazon, M. Shires, D. Crellin, N. Orsi, P. Quirke, and D. Treanor, "Toward routine use of 3D histopathology as a research tool," Amer. J. Pathology, vol. 180, no. 5, 2012.), we improve the performance of registering multistained histology images. Evaluation was conducted on 77 histological image pairs taken from three liver specimens and one intervertebral disc specimen. In total, six types of histochemical stains were tested. We evaluated our method against the same registration method implemented without applying the classification algorithm (intensity-based registration) and the state-of-the-art mutual information based registration. Superior results are obtained with the proposed method. PMID:23955690

  16. Early detection of oral cancer: PAP and AgNOR staining in brush biopsies

    PubMed Central

    Rajput, Dinesh V; Tupkari, Jagdish V

    2010-01-01

    Aim: The aim of this study was to determine the diagnostic accuracy of routine Papanicolaou stain (PAP) and Silver stained Nucleolar Organizer Regions (AgNOR) staining in brush biopsies taken from suspected oral lesions for early detection of oral cancer. Materials and Methods: Brush biopsies were collected from macroscopically suspicious lesions of the oral cavity of 34 patients and 10 normal-aged and sex-matched controls. The numbers of AgNORs were counted in 100 squamous epithelial cell nuclei per slide after silver staining of the smears (Ploton’s one-step method). Results: Sensitivity and specificity of PAP analysis in the oral smears for detection of oral cancer and normal cells was 91.176% and 100%. The positive and negative prediction values were 100% and 76.92%, respectively. Sensitivity and specificity of AgNOR analysis in the oral smears for detection of oral cancer and normal cells was 100%. The positive and negative prediction values were 100% each. Conclusion: Based on the above facts, we conclude that brush biopsy in conjunction with AgNOR staining is an easily practicable, non-invasive, safe and accurate screening method for the detection of macroscopically suspicious oral cancerous lesions. Because of its simple technique and high reliability for cellular proliferation, AgNOR staining in brush smears can be used as an adjunct to other routine cytological diagnoses for the early detection of oral cancer. However, further investigations with more number of study samples will be needed to establish this correlation beyond doubt. PMID:21731263

  17. Peroxisome proliferator-activated receptor-? staining is associated with worse outcome in colorectal liver metastases

    PubMed Central

    PANG, TONY; KAUFMAN, ANTONY; CHOI, JULIAN; GILL, ANTHONY; DRUMMOND, MARTIN; HUGH, THOMAS; SAMRA, JASWINDER

    2015-01-01

    Peroxisome proliferator-activated receptors (PPARs) are a family of nuclear receptors involved in lipid metabolism and liver response to injury. We hypothesised that differences in the expression of PPARs may reflect differences in the cellular microenvironment of the liver and, consequently, in the behaviour of colorectal liver metastases. Of the 145 patients who underwent hepatectomy for colorectal liver metastases between 1998 and 2007, 103 had adequate tissue for PPAR staining and histological re-evaluation. The histological characteristics evaluated included sinusoidal dilatation, perisinusoidal fibrosis, ballooning and steatosis. PPAR- ? and-? staining was performed and the results were correlated with clinical and survival data. Lobular inflammation and sinusoidal dilatation were the most common histopathological abnormalities. A total of 50% of the patients were PPAR- ?-negative and 34% were PPAR- ?-negative. More patients exhibited lobular inflammation in the PPAR- ? -positive group (P=0.023) compared to patients with negative PPAR- ? staining, as seen on the multivariate analysis. PPAR- ?positivity was associated with oxaliplatin use, surgical margins ?1 mm and a trend towards a lesser degree of fibrosis. The median follow-up in this cohort of patients was 48 months. Patients with PPAR- ? staining had a worse overall survival (median, 36 vs. 79 months, P=0.037) compared to those with no PPAR- ? staining. There was no correlation between PPAR- ? or-?positivity and disease-free survival. In conclusion, PPAR- ? staining is associated with lobular inflammation and worse overall survival in patients with colorectal liver metastases. The exact mechanism underlying this finding remains unclear and further research into the diagnostic and therapeutic implications is required. PMID:25798259

  18. A nonlinear mapping approach to stain normalization in digital histopathology images using image-specific color deconvolution.

    PubMed

    Khan, Adnan Mujahid; Rajpoot, Nasir; Treanor, Darren; Magee, Derek

    2014-06-01

    Histopathology diagnosis is based on visual examination of the morphology of histological sections under a microscope. With the increasing popularity of digital slide scanners, decision support systems based on the analysis of digital pathology images are in high demand. However, computerized decision support systems are fraught with problems that stem from color variations in tissue appearance due to variation in tissue preparation, variation in stain reactivity from different manufacturers/batches, user or protocol variation, and the use of scanners from different manufacturers. In this paper, we present a novel approach to stain normalization in histopathology images. The method is based on nonlinear mapping of a source image to a target image using a representation derived from color deconvolution. Color deconvolution is a method to obtain stain concentration values when the stain matrix, describing how the color is affected by the stain concentration, is given. Rather than relying on standard stain matrices, which may be inappropriate for a given image, we propose the use of a color-based classifier that incorporates a novel stain color descriptor to calculate image-specific stain matrix. In order to demonstrate the efficacy of the proposed stain matrix estimation and stain normalization methods, they are applied to the problem of tumor segmentation in breast histopathology images. The experimental results suggest that the paradigm of color normalization, as a preprocessing step, can significantly help histological image analysis algorithms to demonstrate stable performance which is insensitive to imaging conditions in general and scanner variations in particular. PMID:24845283

  19. Ultrasensitive staining-free protein detection after PAA gel electrophoresis using deep UV fluorescence.

    PubMed

    Riaplov, Eugene; Li, Qiang; Seeger, Stefan

    2007-01-01

    We present the observation of separated protein bands after polyacrylamide (PAA) gel electrophoresis based on the staining-free detection of their ultra violet (UV)-induced fluorescence employing deep UV confocal fluorescence microscopy. Mixtures of the three biological compounds beta-Galactosidase (from Escherichia coli), apo-Transferrin (bovine) and bovine serum albumin (BSA) have been separated and a staining free detection limit below 80 pg (7.0 x 10(8) molecules) per band has been achieved. This corresponds to approximately 270 molecules in the detection volume for confocal microscopy. PMID:17897099

  20. [Morphology of low-velocity impact stains produced from single drops of blood].

    PubMed

    Benecke, Mark; Reibe, Saskia; Baumjohann, Kristina; Gulinski, Sarah; Wetzel, Waltraud; Schmidt, Kira; Pressler, Katharina; Lebküchner, Isabell; Streckenbach, Markus

    2012-01-01

    Systematic variation of blood droplet volume, the distance fallen and the surface (paper, wood, plastics, tiles) led to the conclusion that the size and the shape of the stains ("fingers", satellites) allowed to deduce the distance fallen but only if the actual surface structure was known. We found that detailed photography at the crime scene was necessary, yet experiments have to be performed due to the extreme influence of the actual surface texture on all characteristics (size, spines, peripheral spatter) of the blood stains. PMID:22924278

  1. A multiwell plate procedure for immunohistochemical and histochemical staining of constituents of articular cartilage.

    PubMed

    Müller, G; Altenburg, E

    1996-07-01

    A multiwell plate procedure was tested for its applicability to determine immunohistochemically the noncollagenous matrix protein COMP (cartilage oligomeric matrix protein) and histochemically the proteoglycans of the matrix of articular cartilage. Fixed and decalcified cartilage-bone sections were treated with buffer, antisera, substrate or staining solutions in the wells of an assay plate under shaking on a rocking table. This floating procedure results in a reproducible histochemical or immunohistochemical staining and might therefore be valuable to determine or to detect other constituents of the matrix of these connective tissues under comparable conditions. PMID:8863862

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

    PubMed Central

    Watson, Michael L.; Aldridge, William G.

    1961-01-01

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

  3. Gram Stain

    MedlinePLUS

    ... can cause skin infections or pneumonia (also a bioterrorism agent ); Listeria monocytogenes can cause foodborne illnesses . Gram- ... was last modified on February 24, 2015. The review date indicates when the article was last reviewed ...

  4. Silver staining method for DNA in polyacrylamide gels using eriochrome black T as a silver-ion sensitizer.

    PubMed

    Hwang, Sun-young; Jin, Li-tai; Yoo, Gyurng-soo; Choi, Jung-Kap

    2006-05-01

    A sensitive silver staining method using eriochrome black T as a silver-ion sensitizer for DNA detection in polyacrylamide gels was developed. The sensitivity of this staining method was significantly improved by the new silver-ion sensitizer containing a diazo group, which has reducing power. The staining method lasted a total of approximately 15 min following a fixing step for 2 x 20 min. The detection limit of this staining method was 1-4 pg for PhiX174 DNA/HaeIII in both nondenaturing and denaturing polyacrylamide gels. This staining method was especially effective in low-base pair DNA, with a sensitivity that was approximately ten-fold higher than previously published silver staining methods. PMID:16568502

  5. A monoclonal antibody that recognizes a phosphorylated epitope stains lampbrush chromosome loops and small granules in the amphibian germinal vesicle

    PubMed Central

    1990-01-01

    An mAb library was produced against proteins from the germinal vesicle (GV) of the frog Xenopus laevis; mAb 104 was selected from this library on the basis of its immunofluorescent staining of lampbrush chromosome loops. Chromosomes from several species of frogs and salamanders stained equally well. The antibody also stained the surface of numerous small granules in the GV nucleoplasm. The interior of the same granules was stained by antibodies against small nuclear ribonucleoproteins (snRNPs). mAb 104 also stained somatic nuclei from many vertebrate and invertebrate species, usually in a finely punctate pattern similar to that described for anti-snRNP and other antinuclear antibodies. The staining of somatic nuclei was much stronger during the mitotic stages than during interphase. Immunoblot analysis showed that mAb 104 recognizes a phosphorylated epitope. PMID:1703534

  6. Simultaneous determination of l-dopa and its prodrug (S)-4-(2-acetamido-3-ethoxy-3-oxopropyl)-1,2-phenylene diacetate in rat plasma by high-performance liquid chromatography–tandem mass spectrometry and its application in a pharmacokinetic study

    Microsoft Academic Search

    Weizhe Jiang; Li Lv; Songyu Zhou; Xingzhen Huang; Xiaoxia Shi; Cong Lv; Lingling Wu; Chongyao Xu

    2010-01-01

    A sensitive, simple and rapid HPLC–MS\\/MS method has been developed and validated for the simultaneous determination of l-dopa and its prodrug (S)-4-(2-acetamido-3-ethoxy-3-oxopropyl)-1,2-phenylene diacetate (AEPD) in rat plasma in the present study. The analytes were separated on a C18 column (5?m, 2.1mm×150mm) with a security guard C18 column (5?m, 4mm×20mm) and a triple-quadrupole mass spectrometry equipped with an electrospray ionization (ESI)

  7. Darkfield illumination improves microscopic detection of metals in Timm's stained tissue

    Microsoft Academic Search

    Erik Baatrup; Christopher J. Frederickson

    1989-01-01

    Summary  Deposits of trace or toxic metals can be quickly identified by light microscopical surveys of tissue sections stained for metals by variants of Timm's silver enhancement method. The present work shows that the small, isolated silver grains that label isolated deposits of metal in tissue are undetectable in brightfield light microscopy but are easily detected in darkfield microscopy. Darkfield illumination

  8. The development of a nucleus staining fluorescent probe for dynamic mitosis imaging in live cells.

    PubMed

    Ghosh, Krishna Kanta; Jeong, Yun-Mi; Kang, Nam-Young; Lee, JungYeol; Si Yan Diana, Wan; Kim, Jun-Young; Yoo, Jaeduk; Kim, Dohee; Kim, Yun Kyung; Chang, Young-Tae

    2015-05-21

    A low-toxicity nucleus staining fluorescent probe, , was developed for real time mitosis imaging in live cells. was identified by unbiased high-throughput imaging-based screening of a new xanthone library (AX). Unlike the conventional Hoechst dye, the low toxicity of allows long term monitoring of cell division over more than one cell cycle. PMID:25960154

  9. Blood stain pattern interpretation in cases of fatal haemorrhage from ruptured varicose veins

    Microsoft Academic Search

    Roger W. Byard; David Veldhoen; Colin Manock; John D. Gilbert

    2007-01-01

    Blood stain patterns from wounds are determined in part by the nature of the injuries, but also by the types of vessels that have been traumatised. Characteristic spray from arterial injury usually results in a fine projected bloodstain pattern, often found at some distance from the victim. In contrast, venous bleeding tends to be under much lower pressure and less

  10. Differential staining of acid glycosaminoglycans (mucopolysaccharides) by Alcian blue in salt solutions

    Microsoft Academic Search

    J. E. Scott; J. Dorling

    1965-01-01

    The application of the “critical electrolyte concentration” (CEC) concept to the differentiation of acidic glycosaminglycans (mucopolysaccharides) is described. Alcian Blue 8GX stains with increasing selectivity as increasing amounts of magnesium chloride are incorporated into the dye solution. Model experiments with pure polyanions, or artifically carboxylated, phosphorylated and sulphated liver sections, showed that binding of dye to carboxylate or phosphate groups

  11. Sputum gram's stain in community-acquired pneumococcal pneumonia. A meta-analysis.

    PubMed Central

    Reed, W W; Byrd, G S; Gates, R H; Howard, R S; Weaver, M J

    1996-01-01

    The usefulness of the sputum Gram's stain is controversial. This meta-analysis was designed to evaluate the sensitivity and specificity of the sputum Gram's stain in community-acquired pneumococcal pneumonia. Using a predetermined protocol, articles were discovered through a MEDLINE search (1966 to 1993) and the examination of bibliographies and were graded for quality by three blinded reviewers. Information on the reference standard, blinding, stain interpreter, control for antibiotic use, and definition of a positive test was collected. We found 12 articles containing 17 test characteristics to evaluate. The number of patients in each study ranged from 16 to 404. Sputum culture was the most common reference standard (10 of 17 estimations). Sensitivity ranged from 15% to 100% and specificity from 11% to 100%. Test characteristics varied markedly among studies and appeared related partly to the test interpreter. The sputum Gram's stain may yield misleading results in community-acquired pneumonia, as its sensitivity and specificity vary substantially in different settings. A practitioner electing to use the study should be well trained and use a specific definition for a positive test. PMID:8987424

  12. Marker-controlled watershed segmentation of nuclei in H&E stained breast cancer biopsy images

    Microsoft Academic Search

    M. Veta; A. Huisman; M. A. Viergever; P. J. van Diest; J. P. W. Pluim

    2011-01-01

    In this paper we present an unsupervised automatic method for segmentation of nuclei in H&E stained breast cancer biopsy images. Colour deconvolution and morphological operations are used to preprocess the images in order to remove irrelevant structures. Candidate nuclei locations, obtained with the fast radial symmetry transform, act as markers for a marker-controlled watershed segmentation. Watershed regions that are unlikely

  13. Genetic mapping of resistance to purple seed stain in PI 80837 soybean

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Purple seed stain (PSS) of soybean caused by Cercospora kikuchii is an important disease that reduces market grade and can affect seed germination and vigor. A single dominant gene was shown to confer PSS resistance in PI 80837. The objective of this research was to map the PSS resistance gene in P...

  14. Detection of firearm discharge residues in blood-stained articles by fluorescence.

    PubMed

    Nag, N K; Mazumdar, M

    1975-02-01

    A quick and sensitive method has been developed for the chemical detection of nitrous derivatives discharged from a firearm. Detection by fluorescence is especially suitable for blood-stained articles such as clothing, which often give confusing results when the conventional methods are used. PMID:1132864

  15. Gradual oxidation of stain etched porous silicon nanostructures applied to silicon-based solar cells

    Microsoft Academic Search

    B. González-Díaz; R. Guerrero-Lemus; J. Méndez-Ramos; B. Díaz-Herrera; V. D. Rodríguez

    2009-01-01

    This work describes the photoluminescence (PL) and morphology of stain etched porous silicon nanostructures (PSN) submitted to a gradual oxidation by immersion in a HNO3 boiling point solution and applied to solar cells. The gradual oxidation passivates the extremely reactant fresh porous surface and softens the porous structure for an adequate placement of the metallic contact on top. The pore

  16. Application of Stain Etched Porous Silicon in Solar Cells and Light Emitting Diodes

    Microsoft Academic Search

    D. Dimova-Malinovska

    A review of the properties of porous silicon (PS), prepared by stain etching, and of light emitting and solar cell heterostructures\\u000a utilising it is presented. The mechanisms of carrier transport in light emitting diode (LED) structures and of the electroluminescence\\u000a are also discussed.

  17. Changes in absorption, fluorescence, dichroism, and birefringence in stained giant axons: Optical measurement of membrane potential

    Microsoft Academic Search

    W. N. Ross; B. M. Salzberg; L. B. Cohen; A. Grinvald; H. V. Davila; A. S. Waggoner; C. H. Wang

    1977-01-01

    Summary The absorption, fluorescence, dichroism, and birefringence of stained squid axons were measured during action potentials and voltage clamp steps in an effort to find large optical signals that could be used to monitor membrane potential. Changes in all four optical properties were found that were linearly related to membrane potential and, with several new dyes, the signal-to-noise ratios were

  18. A novel medical robot assisting photodynamic therapy for port wine stains

    Microsoft Academic Search

    Gui-bin Bian; Xing-guang Duan; Qiang Huang; Xing-tao Wang; Shi-hu Cui; Naiyan Huang; Ying Gu

    2010-01-01

    Port wine stain (PWS) birthmarks are congenital vascular malformations, which usually appear at birth and tend to become darker and thicker with age's growth. Vasculartargeted photodynamic therapy (PDT) is an effective approach in the treatment of PWS. However, due to the arbitrariness of manual operation and pole points existing in laser radiation, the PWS zone was always cured unevenly in

  19. Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections

    Microsoft Academic Search

    L. C. U. Junqueira; G. Bignolas; R. R. Brentani

    1979-01-01

    Synopsis  Sirius Red, a strong anionic dye, stains collagen by reacting, via its sulphonic acid groups, with basic groups present in the collagen molecule. The elongated dye molecules are attached to the collagen fibre in such a way that their long axes are parallel. This parallel relationship between dye and collagen results in an enhanced birefringency.Examination of tissue sections from 15

  20. Design Implications from a Usability Study of GramStain-Tutor.

    ERIC Educational Resources Information Center

    Kim, Sara; Brock, Douglas; Orkand, Adam; Astion, Michael

    2001-01-01

    Describes a usability study conducted with health sciences students at the University of Washington that explored interface issues in the GramStain Tutor, an educational software program on CD-ROM, particularly the navigation of the program and the use of embedded design features. (LRW)

  1. The potential of the immunogold-silver staining method for paraffin sections

    Microsoft Academic Search

    D. R. Springall; G. W. Hacker; L. Grimelius; J. M. Polak

    1984-01-01

    The immunogold-silver staining technique is shown to be of great value in the detection of regulatory peptide-containing nerves and endocrine cells in routinely fixed, paraffin-wax-embedded tissues. The method appears to be better for this system than peroxidase anti-peroxidase (PAP) which can yield poor or variable results.

  2. Fungal proteins with mannanase activity identified directly from a Congo Red stained zymogram by mass spectrometry.

    PubMed

    Peterson, Robyn; Grinyer, Jasmine; Joss, Janice; Khan, Alamgir; Nevalainen, Helena

    2009-12-01

    Secreted fungal proteins with mannanase activity were identified by mass spectrometry of bands excised from a Congo Red stained zymogram containing locust bean gum as substrate. This technique circumvents the need to locate corresponding bands on a parallel gel without substrate and provides good accuracy in targeting proteins for identification. PMID:19854225

  3. New staining methods for sperm evaluation estimated by microscopy and flow cytometry

    Microsoft Academic Search

    M. Magistrini; E. Guitton; Y. Levern; J. Cl. Nicolle; M. Vidament; D. Kerboeuf; E. Palmer

    1997-01-01

    New staining methods and automated instruments are now available to evaluate the sperm cell in vitro. Individual compartments of the sperm cell, such as the nucleus and the plasma and acrosomal membranes, may be investigated, as well as the cell function as shown by mitochondria activity and capacitation. Various probes are used and they can be analyzed by direct light

  4. Quick histochemical staining method for measuring lactate dehydrogenase C4 activity in human spermatozoa.

    PubMed

    Cui, Zhaolei; Chen, Liangyuan; Liu, Yaohua; Zeng, Zhangxin; Lan, Fenghua

    2015-04-01

    The enzyme activity of lactate dehydrogenase C4 (LDH-C4, due to tetrameric nature of C-subunit) has been proposed as an important parameter in evaluating sperm motility and semen quality. A novel histochemical staining method for detecting LDH-C4 activity in human spermatozoa is described in this report. The staining working solution comprises sodium 2-hydroxybutyrate (an affinity substrate of LDH-C4), nitrotetrazolium blue chloride (NBT), nicotinamide adenine dinucleotide (NAD) and naphthol blue. The positive products were purple black lumps concentrated in the neck segment of the spermatozoa and weakly in the middle piece. A normal reference range for the integral enzyme activity was constructed from 120 healthy males based upon the scoring criteria. The study further compared the staining method with the routine spectrophotometry technique in terms of the results of 96 cases with infertile status. Moreover, we found the down-regulated LDH-C4 expression was significantly correlated with the lowered enzyme activity (r=0.865, P=0.000). Our data suggest that the histochemical staining method hallmarks a relatively high accuracy and may be a better alternative for measuring LDH-C4 activity in human spermatozoa. PMID:25795631

  5. THE CYTOCHEMICAL STAINING AND MEASUREMENT OF PROTEIN WITH MERCURIC BROMPHENOL BLUE

    Microsoft Academic Search

    PHILIP A. BREWER; MAX ALFERT

    The development of chromatographic and electrophoretic techniques which re- quire identification of spots on filter paper has given new impetus to the study of color reactions of amino acids and proteins, and it is inevitable that the experience which is developing rapidly in this field will be carried over to the field of biological staining. Durruln (1950) devised the mercuric

  6. Blood flow dynamics after laser therapy of port wine stain birthmarks

    Microsoft Academic Search

    Yu-Chih Huang; Nadia Tran; E. Victor Ross; Peter R. Shumaker; J. Stuart Nelson; Kristen Kelly; Bernard Choi

    2009-01-01

    During laser therapy of port wine stain (PWS) birthmarks, regions of persistent perfusion may exist. We hypothesize that such regions, which are not readily visible, exist even during laser surgery performed by highly experienced clinicians. The objective of this study was to use objective feedback to assess the acute vascular response to laser therapy. We have developed a clinic-friendly laser

  7. Brilliance All Around: The Stained Glass of Sterling and Its Maker

    E-print Network

    Crair, Michael C

    of students, staff, and visitors, these images in glass deserve close attention, and their maker, G. Owen to carry out extensive research into the images, the artist, stained glass in the United States, American noticed that the lunch room window showing Jack Spratt and his wife displayed leaded windows

  8. Improved selective, simple, and contrast staining of acidophilic neurons with vanadium acid fuchsin

    Microsoft Academic Search

    Ilya V Victorov; Konstantin Prass; Ulrich Dirnagl

    2000-01-01

    Acidophilia is one of the hallmarks of acute neuronal damage and death in brain ischemia, excitotoxic and traumatic lesions and epileptic seizures. We here describe a novel and simple method for visualizing acidophilic neurons on paraffin sections, using vanadium acid fuchsin (VAF) staining and toluidine blue or hematoxylin counterstaining. Paraffin sections of the brain fixed in ethanol–formalin–acetic acid mixture are

  9. Building Partnership: Gail M. Staines--Western NY Library Resources Council, Buffalo

    ERIC Educational Resources Information Center

    Library Journal, 2004

    2004-01-01

    This article details the work of Gail Staines, who is probably the only Mover & Shaker who lists the U.S.A. Equestrians Association among her professional memberships. But it is not that far a leap from what she does in her work as director of the Western New York Library Resources Council (WNYLRC), where she is responsible for making 98 libraries…

  10. Heterogeneous staining: a tool for studies of how fluorescent dyes affect the physical properties of DNA

    PubMed Central

    Nyberg, Lena; Persson, Fredrik; Åkerman, Björn; Westerlund, Fredrik

    2013-01-01

    The commonly used fluorescent dye YOYO-1 (YOYO) has, using bulk techniques, been demonstrated to stain DNA heterogeneously at substoichiometric concentrations. We here, using nanofluidic channels and fluorescence microscopy, investigate the heterogeneous staining on the single DNA molecule level and demonstrate that the dye distribution is continuous. The equilibration of YOYO on DNA is extremely slow but can be accelerated by increasing the ionic strength and/or the temperature. Furthermore, we demonstrate how to use the heterogeneous staining as a tool for detailed and time-efficient studies of how fluorescent dyes affect the physical properties of DNA. We show that the relative increase in extension of DNA with increasing amount of YOYO bound is higher at low ionic strengths and also extrapolate the extension of native DNA. Our study reveals important information on how YOYO affects the physical properties of DNA, but it also has broader applications. First, it reveals how cationic intercalators, such as potential DNA drugs, affect DNA under strong confinement. Second, the strategy of using heterogeneous staining is of general use for single molecule studies of DNA interacting with proteins or ligands. PMID:23975199

  11. Remarks on the usefulness of toluidine blue staining for RNA cytophotometry in plastic embedded tissues.

    PubMed

    Meyer, W; Zschemisch, N H

    1999-06-01

    The study demonstrates the usefulness of water-soluble plastic resins for the cytological quantification of RNA contents after toluidine blue staining. In this way shrinkage artefacts in delicate tissues are avoided and more exact cytophotometrical results can be obtained from embryological material. PMID:10432183

  12. CHARACTERIZATION OF ORGANIC EMISSIONS FROM A WOOD FINISHING PRODUCT - WOOD STAIN

    EPA Science Inventory

    The paper gives results of the measurement of emission characteristics of four organic compounds (nonane, decane, undecane, and 1,2,4-trimethylbenzene) from a wood finishing product, wood stain, in an environmental chamber. It was found that the emission patterns of the four orga...

  13. Meconium stained amniotic fluid in preterm delivery is an independent risk factor for perinatal complications

    Microsoft Academic Search

    Moshe Mazor; Reli Hershkovitz; Asher Bashiri; Eli Maymon; Ruth Schreiber; Doron Dukler; Miriam Katz; Ilana Shoham-Vardi

    1998-01-01

    Objective: To determine the prevalence and clinical significance of meconium stained amniotic fluid (MSAF) in women with preterm delivery. Study design: The study population consisted of consecutive patients who arrived with intact membranes and delivered preterm, singleton neonates at the Soroka Medical Center between 1 January 1985 and 31 December 1995. Only vertex presentation was included. Antepartum death was excluded

  14. Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopy

    PubMed Central

    Vogel, Benjamin; Siebert, Hanna; Hofmann, Ulrich; Frantz, Stefan

    2015-01-01

    Picrosirius red (PSR) staining is a commonly used histological technique to visualize collagen in paraffin-embedded tissue sections. PSR stained collagen appears red in light microscopy. However it is largely unknown that PSR stained collagen also shows a red fluorescence, whereas live cells have a distinct green autofluorescence. Both emission patterns can be detected using standard filter sets as found in conventional fluorescence microscopes. Here we used digital image addition and subtraction to determine the relative area of the pure collagen and live cell content in heart tissue in a semi-automated process using standard software. This procedure, which considers empty spaces (holes) within the section, can be easily adapted to quantify the collagen and live cell areas in healthy or fibrotic tissues as aorta, lung, kidney or liver by semi-automated planimetry exemplified herein for infarcted heart tissue obtained from the mouse myocardial infarction model. • Use of conventional PSR stained paraffin-embedded tissue sections for fluorescence analysis. • PSR and autofluorescence images are used to calculate area of collagen and area of live cells in the tissue; empty spaces (holes) in tissue are considered. • High throughput analysis of collagen and live cell content in tissue for statistical purposes.

  15. Image staining and differential diagnosis of ultrasound scans based on the Mahalanobis distance

    Microsoft Academic Search

    Reza Momenan; Robert F. Wagner; Brian S. Garra; Murray H. Loew; Michael F. Insana

    1994-01-01

    The covariance matrices associated with each state of health or disease from a previous study are used as the basis of an image staining display technique for aid in quantitative differential diagnosis. A state of health or disease is chosen by the clinician: this selects the covariance matrix from the data base. A region of interest (ROI) is then scrolled

  16. The choice of a masking agent in the histochemical staining of metals

    Microsoft Academic Search

    Y. Sumi; T. Muraki; T. Suzuki

    1983-01-01

    Summary  The masking effects of standard masking agents (aminopolycarboxylic acids, carboxylic acids and phosphates) have been investigated in both test-tube experiments and tissue sections in order to ascertain the factors which must be considered when choosing a masking agent for the histochemical staining of a metal. The masking effectsin vitro were determined by spectrophotometry through the complexing of the dye Chrome

  17. Detecting microcalcifications in atherosclerotic plaques by a simple trichromic staining method for epoxy embedded carotid endarterectomies.

    PubMed

    Relucenti, M; Heyn, R; Petruzziello, L; Pugliese, G; Taurino, M; Familiari, G

    2010-01-01

    Atherosclerotic plaques have a high probability of undergoing rapid progression to stenosis, becoming responsible of acute coronary syndrome or stroke. Microcalcifications may act as enhancers of atherosclerotic plaque vulnerability. Considering that calcifications with a diameter smaller than 10 mm in paraffin embedded tissue are rather difficult to detect, our aim was to analyze microcalcifications on semithin sections from epoxy resin embedded samples of carotid endarterectomies using an original trichromic stain (methylene blue--azur B--basic fuchsine--alizarin red). We have compared samples stained either with our method, methylene blue-azur B alone or with Von Kossa staining, and methylene blue-azur B -basic fuchsine alone or with Von Kossa staining. Our method resulted to be simple and fast (ca. 2 min), it gives a sharp general contrast for all structures and allows to easy identify collagen and elastin. In addition, gray-green colour associated to intracellular lipid droplets evidences foam cells, which are particularly abundant in endarterectomies samples. Mast cells and their metachromatic granules are also well recognized. Calcifications over 0,5 mm are clearly recognizable. In conclusion, microcalcifications are clearly distinguished from the extracellular matrix in spite of their reduced dimensions. Methylene blue--azur B--basic fuchsine--alizarin red method is easy to use, reproducible, and is particularly suitable for the identification of microcalcifications in the morphological analysis of atherosclerotic plaques. PMID:20819772

  18. Recruitment of OKM1 staining lymphocytes with selective binding to K-562 tumour targets by interferon.

    PubMed Central

    Salata, R A; Schacter, B Z; Ellner, J J

    1983-01-01

    Spontaneous cytotoxicity of human lymphocytes for tumours is increased by interferon (IFN) without change in the overall fraction of cells binding to targets. We developed an indirect immunofluorescent technique to stain lymphocytes conjugated to K-562 tumour cells in agarose with monoclonal antibodies. This allowed assessment of lymphocyte subpopulations binding to tumour cells without disruption of conjugates. Overall binding of non-adherent (NA) lymphocytes to tumour targets following incubation at 37 degrees C for 6 h was 13.3 +/- 0.3% compared to 12.5 +/- 0.7% with inclusion of IFN at 100 u/ml. When NA lymphocytes were incubated with K-562 tumour cells without IFN, OKM1 and OKT3 staining lymphocytes comprised 16.8 +/- 3.5% and 83.0 +/- 1.3% of the total lymphocyte population and 32.5 +/- 1.3% and 70.2 +/- 2.6% of lymphocytes conjugated to tumours. Incubation with IFN significantly increased OKM1 staining cells in the total NA population to 57.2 +/- 5.6% (P less than 0.01) and within tumour conjugates to 59.2 +/- 2.7% (P less than 0.01) while OKT3 staining cells decreased to 58.3 +/- 5.2% (P less than 0.02) and 45.3 +/- 1.2% (P less than 0.01), respectively. IFN increased cytotoxicity of NA cells for 51Cr-labelled K-562 by 66% at an effector to target ratio of 30:1 (P less than 0.001). These results demonstrate that OKM1 staining cells bind more avidly to tumour targets in the absence of IFN. IFN selectively increases the proportion of OKM1 staining lymphocytes with a concomitant increase in their binding to tumour cells. Therefore, enhancement of cytotoxicity by IFN in the NK system may result, in part, from conversion of OKT3 to OKM1 staining cells which are more efficient killers. PMID:6190593

  19. Detection of Candida albicans in oral squamous cell carcinoma by fluorescence staining technique

    PubMed Central

    Jahanshahi, Gholamreza; Shirani, Samaneh

    2015-01-01

    Background: One of the probable etiologic risk factors of oral squamous cell carcinoma (OSCC) is Candidal infection, especially by Candida albicans, whose role has not definitely been confirmed. Some have assigned a primary role to Candida, whereas others consider it as a transient inhabitant. The debate may be due to lack of an accurate and sensitive revealing technique. By identifying the presence of Candida, especially in deeper parts of OSCC, the etiologic role may be verified. The present study was conducted to detect the presence of Candida in OSCC by fluorescence staining technique. Materials and Methods: This study was descriptive experimental. Calcofluor-white, which is applied in fluorescence staining, is a specific staining substance for Candida and has a higher accuracy compared with other common methods. 100 specimens of well-differentiated OSCC with adequate amount of tissue were retrieved from the archive and two serial sections were obtained from each one. The first section was stained using the popular histochemical (periodic acid-Schiff [PAS]) method and then evaluated under a light microscope to detect the presence of Candida. The second section was stained using fluorescence staining technique. The sum of counted Candida in each technique was fed into SPSS software and analyzed by McNamara test. P < 0.001 was considered as significant. Results: The amount of Candida present in OSCCs was 74% measured by fluorescence technique. The sensitivity and specificity of the two staining techniques were significantly different. These parameters in the fluorescence technique were higher than those of the histochemical (PAS) method, confirmed by McNamara test showing significantly different results for them (P < 0.001). The results obtained from the fluorescence technique had higher accuracy compared with the histochemical (PAS) method. Conclusion: Some researchers couldn’t find a considerable number of fungi in OSCC, while our results revealed more presence of Candida, especially in deeper parts of tissue samples and probably a more important role for Candida as an etiologic risk factor for OSCC. However, since the fluorescence technique had a higher accuracy in the identification of Candida and it was nearly evident in two-third of the samples, the role of fungi as a primary cause is suggested to be studied in future investigations. PMID:25878675

  20. Pyrogallol red-vanadium complex-a new stain for electron microscopy.

    PubMed

    Völker, W; Kampsmeyer, H H; Robenek, H

    1996-11-01

    We report on the application of a pyrogallol red-vanadium complex (PR-V) for ultracytochemical staining of proteinaceous structures in animal tissues and cell cultures. This dye may be used as a general purpose stain in electron microscopy. In contrast to osmium tetroxide, the price of the material is low and no toxic vapors are produced. The PR-V complex was prepared by addition of vanadium (IV) oxide sulfate to pyrogallol red dissolved in acetate buffer (pH 5.6). The formation of the complex was indicated by a color change from purple-red (lambda max = 520 nm) to violet (lambda max = 539 nm) which occurred at equimolar concentrations of the dye and the metal salt. Under these conditions PR-V was stable for several days. The mechanism of PR-V binding was checked in dot blots using different proteins as well as heparin for control. While heparin remained unstained, proteins were stained in a dose-dependent manner. Deamination of proteins with nitric oxide strongly reduced PR-V staining in dot blots as well as in cell cultures. Optimal staining results of animal cells and tissues were obtained in specimens that had been mildly fixed for at least 1 h or longer with a mixture of 0.1% glutaraldehyde and 1.0% paraformaldehyde dissolved in phosphate-buffered saline, pH 7.2, washed with acetate buffer, pH 5.6, and subsequently treated with PR-V in the presence of 50% ethanol at room temperature. Control specimens without PR-V but treated en bloc with uranyl acetate or sodium molybdate showed similar contrast but less details in the ultrastructure of the tissue. All specimens were embedded in epoxy resin and ultratain sections were stained conventionally with uranyl and lead salt solutions. In electron micrographs, membrane-associated particles, stress fibers and filaments of the cell cortex, collagen fibrils, tight junctions and desmosomes, and other proteinaceous components were clearly visualized only in the PR-V-treated specimens. In conclusion, the ability to bind selectively and specifically to protein-aceous structures makes PR-V a versatile stain to study the localization and distribution of these structures in cells and tissues at the ultrastructural level. PMID:8950609

  1. Histamine immunohistochemistry is superior to the conventional heparin-based routine staining methodology for investigations of human skin mast cells.

    PubMed

    Johansson, O; Virtanen, M; Hilliges, M; Yang, Q

    1994-05-01

    Conventional studies of mast cells are limited by methodological restrictions such as a selective fixative-dependent routine staining blockage. This is thought to depend on the biochemical differences of the mast cell granule contents suggesting a cellular heterogeneity. Investigations of human mast cells, using routine methods, also suffer from the problem of a low signal-to-noise ratio. In the present study, normal human skin was used to compare an immunohistochemical method for histamine with two recommended mast-cell fixatives and a new commercial fixative in combination with three routine stains. Mast cells were found throughout the dermis with all the routine stains used. However, immunohistochemistry gave profoundly better results. Small structures, such as thin cytoplasmatic extensions and single granules, were readily detectable. Double-staining (immunohistochemistry followed by routine staining) revealed differences in staining capacity. All immunoreactive cells were not stained by routine stains and sometimes the opposite was also seen. This supports earlier reported evidence of heterogeneity, not only between skin and intestinal mast cells but also among skin mast cells themselves. Furthermore, by focusing on histamine, instead of heparin, we probably overcame the problems of the selective fixative-dependent routine staining blockage. Finally, the immunofluorescence technique provides a high signal-to-noise ratio and is an excellent method for making high-quality microphotographs of human mast cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8045782

  2. Rapid and easy protein staining for SDS-PAGE using an intramolecular charge transfer-based fluorescent reagent.

    PubMed

    Suzuki, Yoshio; Yokoyama, Kenji; Namatame, Ichiji

    2006-09-01

    High-performance staining for 1-D and 2-D SDS-PAGE was carried out using a novel protein-binding fluorophore (Dye 1), which noncovalently interacts with proteins and provides a fluorescence emission response to proteins by intramolecular charge transfer. In order to achieve the high-throughput analysis of proteins for SDS-PAGE, the general protocols for in-gel protein staining (SDS-PAGE, fixation, staining, washing, and detection) were simplified to produce an easy and rapid protocol (SDS-PAGE together with staining, washing, and detection). This method was performed by preparation of an electrophoresis buffer containing Dye 1 under optimum conditions, and by the binding of Dye 1 to proteins in the gel during the SDS-PAGE. As a result, this study required only 15 min for protein staining as a minimum time. On the other hand, it takes several hours for the general protein staining method, such as SYPRO Ruby staining (18 h) and CBB staining (105 min). Moreover, the protein-to-protein variation was low, and the detection limit was 7.0 ng/band of BSA (S/N = 3.0) in this method, which was as sensitive as the short-protocol silver staining methods. On the basis of these results, this rapid and easy protocol for SDS-PAGE using Dye 1 may be widely applicable and convenient for users in the various scientific and medical fields. PMID:16944465

  3. Mast cells of the bovine trachea: staining characteristics, dispersion techniques and response to secretagogues.

    PubMed Central

    Harris, W H; Marshall, J S; Yamashiro, S; Shaikh, N

    1999-01-01

    Sections of the lower trachea of cattle, fixed in either Carnoy's or formalin, were stained with toluidine blue, alcian blue, or alcian blue and safranin O to study the mast cell population. After toluidine blue staining, about twice as many cells in tissue fixed in Carnoy's contained dark blue granules compared with tissue fixed in formalin. In addition, for the first time in cattle, a population of cells containing red granules was identified after staining with alcian blue and safranin O. Most of these red granules were formalin sensitive. An enzymatic dispersal technique for mast cells is described that yielded 9.4+/-0.4% mast cells (percentage of nucleated cells) with a viability of 92.3+/-0.6%. Spontaneous histamine release was 3.3+/-0.8%. Dispersed mast cells were challenged with various immunological and nonimmunological secretagogues. The calcium ionophores, A23187, ionomyocin, and BrX537A, were effective in releasing up to 94% of histamine in mast cells in a dose-response relationship. Pasteurella haemolytica culture supernate caused about 10% histamine release at a dose of 0.5 mg/mL after correction for spontaneous release. The average histamine content of the mast cells was 6.6+/-1.0 pg/cell. Cytospins of dispersed cells fixed in Carnoy's and stained with alcian blue and safranin O contained mast cells with blue and red granules, and a few cells with a mixture of both granule types. Based on the effects of type of fixation, staining characteristics and histamine content, a mix of subtypes of mast cells is present in the bovine trachea. However, functionally they respond to secretagogues differently than rodent mast cells. Without an immunological secretagogue, studies to determine compounds that will be effective in blocking mast cell degranulation will be limited. Images Figure 2. PMID:9918327

  4. In vivo and in vitro staining of acidophilic neurons as indicative of cell death following kainic acid-induced lesions in rat brain

    Microsoft Academic Search

    G. J. Lees

    1989-01-01

    An in vivo method for positively staining dead neurons was developed and compared with an in vitro staining method using acid fuchsin. Neurons previously killed by intracerebral injections of kainic acid were selectively stained by trypan blue within 15 min of its injection in vivo into the central nervous system of rats. Such staining persisted for at least 4 days

  5. Detection of radioactively labeled proteins is quenched by silver staining methods: quenching is minimal for ¹⁴C and partially reversible for ³H with a photochemical stain

    Microsoft Academic Search

    M. L. Van Keuren; D. Goldman; C. R. Merril

    1981-01-01

    Silver staining methods for protein detection in polyacrylamide gels have a quenching effect on autoradiography and fluorography. This effect was quantitated for proteins in two-dimensional gels by microdensitometry using a computer equipped with an image processor and by scintillation counting of proteins solubilized from the gels. The original histologically derived silver stain had a quenching effect that was severe and

  6. Detection of radioactively labeled proteins is quenched by silver staining methods: quenching is minimal for /sup 14/C and partially reversible for /sup 3/H with a photochemical stain

    SciTech Connect

    Van Keuren, M.L.; Goldman, D.; Merril, C.R.

    1981-09-15

    Silver staining methods for protein detection in polyacrylamide gels have a quenching effect on autoradiography and fluorography. This effect was quantitated for proteins in two-dimensional gels by microdensitometry using a computer equipped with an image processor and by scintillation counting of proteins solubilized from the gels. The original histologically derived silver stain had a quenching effect that was severe and irreversible for /sup 3/H detection and moderate for /sup 14/C detection. A silver stain based on photochemical methods had minimal quenching of /sup 14/C detection and less of a quenching effect than the histological stain for /sup 3/H detection. The /sup 3/H quenching effect was partially reversible for the photochemical stain.

  7. A novel method of combining Periodic Acid Schiff staining with Wright-Giemsa staining to identify the pathogens Penicillium marneffei, Histoplasma capsulatum, Mucor and Leishmania donovani in bone marrow smears

    PubMed Central

    QIN, LINGYAN; ZHAO, LIGANG; TAN, CHUNYAN; CHEN, XU; YANG, ZHENG; MO, WUNING

    2015-01-01

    Penicillium marneffei, Histoplasma capsulatum, Mucor and Leishmania donovani can lead to penicilliosis marneffei, histoplasmosis, mucormycosis and leishmaniasis, respectively, which, to a certain extent, share similar clinical manifestations. These pathogens are approximately the same size, therefore it is relatively difficult to rapidly diagnose the diseases. The aim of the present study was to explore a novel method that attempts to rapidly identify the pathogens of these diseases. In the Wright-Giemsa staining, the four pathogens were approximately the same size and mainly existed in macrophages. The multiplying P. marneffei had two nuclei, which were on both sides of the fungus, and had light cross-walls in the middle. H. capsulatum had a purplish nucleus, which occupied between one-third and one-half of the spore. The cytoplasm was light blue. Peripheral spores were observed in the form of an empty, bright ring without color, like a capsule. Generally, Mucor were observed to have a long and lightly stained area, which could be easily confused with the Wright staining of dinuclear P. marneffei. L. donovani exhibited a deep-staining kinetoplast near the nucleus. In the Periodic Acid Schiff (PAS) staining, the pathogens of P. marneffei and H. capsulatum were distinct and stained red. Differentiation between P. marneffei and H. capsulatum relied on their modes of reproduction: P. marneffei depends on fission, when the pathogens stretch into sausage-shapes and are split by a cross-wall, while H. capsulatum depends on budding so that narrow-necked, single spores can be formed. With PAS staining, the cell walls and intracellular contents of Mucor and L. donovani were not stained, lightly stained or granulated and discontinuous. In conclusion, this method, combining PAS and Wright-Giemsa staining, is simple and rapid, and may contribute to the effective identification of the four pathogens.

  8. Comparative analysis of H&E and Prussian blue staining in a mouse model of cerebral microbleeds.

    PubMed

    Liu, Shuo; Grigoryan, Mher Mahoney; Vasilevko, Vitaly; Sumbria, Rachita K; Paganini-Hill, Annlia; Cribbs, David H; Fisher, Mark J

    2014-11-01

    Cerebral microbleeds are microscopic hemorrhages with deposits of blood products in the brain, which can be visualized with MRI and are implicated in cerebrovascular diseases. Hematoxylin and eosin (H&E) and Perl's Prussian blue are popular staining methods used to localize cerebral microbleeds in pathology. This paper compared these two staining techniques in a mouse model of cerebral microbleeds. We used lipopolysaccharide (LPS) to induce cerebral microhemorrhages. C57B6 mice were treated with LPS (5 mg/kg, i.p.) or vehicle at baseline and at 24 hr. The brains were extracted 48 hr after the first injection and adjacent coronal sections were stained with H&E and Prussian blue to compare the effectiveness of the two staining techniques. H&E-positive stains were increased with LPS treatment and were correlated with grossly visible microhemorrhages on the brain surface; Prussian blue-positive stains, by comparison, showed no significant increase with LPS treatment and did not correlate with either H&E-positive stains or surface microhemorrhages. H&E staining is thus a more reliable indicator of acute bleeding events induced by LPS in this model within a short time span. PMID:25063000

  9. ROBUST METAL COMPLEXES, FERROCENYLMETHYL CARBOXYHYDRAZIDE AND 1-CHLOROMERCURIFERROCENE, AS ELECTRON-OPAQUE STAINS FOR ALDEHYDES AND THIOL GROUPS

    Microsoft Academic Search

    DAVID E. ALLEN; DOUGLAS D. PERRIN

    Ferrocenylmethyl carboxyhydrazide has been used for the ultrastructural localization of glycogen, the glycocalyx of cat intestine and deoxyribonucleic acid. l-Chloromercurifer- rocene is introduced as a stain for thiol groups in wool. Both compounds are examples of a potentially useful class of cytochemical stains made from a \\

  10. Carbon and oxygen isotope geochemistry of live (stained) benthic foraminifera from the Aleutian Margin and the Southern Australian Margin

    E-print Network

    Levin, Lisa

    Carbon and oxygen isotope geochemistry of live (stained) benthic foraminifera from the Aleutian October 2008 Accepted 4 November 2008 Keywords: stable isotopes benthic foraminifera 13C 18O deep sea-water geochemistry and stable isotopic values of the tests of living (stained) calcareous benthic foraminifera from

  11. Association between Black Stains and Dental Caries in Primary Teeth: Findings from a Brazilian Population-Based Birth Cohort

    PubMed Central

    França-Pinto, C.C.; Cenci, M.S.; Correa, M.B.; Romano, A.R.; Peres, M.A.; Peres, K.G.; Matijasevich, A.; Santos, I.S.; Barros, A.J.D.; Demarco, F.F.

    2012-01-01

    Lower dental caries experience has been observed in children and teenagers with the presence of black stains on dental structures. However, none of the previous investigations were population-based studies or adjusted the analysis for potential confounders. This study assessed the prevalence of black stains at the age of 5 in a population-based birth cohort from Pelotas, Brazil and investigated the association between black stains and dental caries. A total of 1,129 children from the 2004 Pelotas birth cohort were examined at age 5, and their mothers were interviewed at their households. Dental examinations included a search for black stains and dental caries on the primary dentition through the dmf-s index. The mothers’ questionnaire comprised data on demographic, social, and behavior aspects. Prevalence of black stains was 3.5% (95% CI 2.5–4.7) and the prevalence of dental caries was 48.4% (95% CI 45.4–51.4). Multivariable logistic regression analysis was performed to assess the association between black stains and dental caries. Adjusted analysis revealed that the presence of black stains was associated with lower levels of dental caries (OR = 0.51; 95% CI 0.26–0.99). The results of the present study suggest that black stains are a protective factor for dental caries development. PMID:22488298

  12. Order-to-Disorder Transition in Ring-Shaped Colloidal Stains A lvaro G. Marin,* Hanneke Gelderblom,

    E-print Network

    Snoeijer, Jacco

    Order-to-Disorder Transition in Ring-Shaped Colloidal Stains A´ lvaro G. Mari´n,* Hanneke Gelderblom, Detlef Lohse, and Jacco H. Snoeijer§ Physics of Fluids Group, Faculty of Science and Technology reveal a structural transition in the stain, from ordered crystals to disordered packings. We show

  13. Effects of molecular oxygen on detection of superoxide radical with nitroblue tetrazolium and on activity stains for catalase

    Microsoft Academic Search

    D. A. Clare; M. N. Duong; D. Darr; F. Archibald; I. Fridovich

    1984-01-01

    The usual method of staining polyacrylamide gel electropherograms for superoxide dismutase activity utilizes a photochemical flux of Oâ⁻ to reduce nitroblue tetrazolium. Superoxide dismutases intercept Oâ⁻, preventing formazan production and thus causing achromatic bands. In the presence of HâOâ, catalase also yield achromatic bands during this staining procedure. This is due to local elevation of pOâ by the catalytic decomposition

  14. Colloidal Silver Staining of Electroblotted Proteins for High Sensitivity Peptide Mapping by Liquid Chromatography–Electrospray Ionization Tandem Mass Spectrometry

    Microsoft Academic Search

    Inge van Oostveen; Axel Ducret; Ruedi Aebersold

    1997-01-01

    Mass spectrometric techniques for the identification of proteins either by amino acid sequencing or by correlation of mass spectral data with sequence databases are becoming increasingly sensitive and are rapidly approaching the limit of detection achieved by the staining of proteins in gels or, after electroblotting, on membranes. Here we present a technique for the sensitive staining of proteins electroblotted

  15. Research article Using the new Phadebas1 Forensic Press test to find crime scene saliva stains suitable for DNA analysis

    Microsoft Academic Search

    Johannes Hedman; Karin Gustavsson; Ricky Ansell

    The Phadebas1 Forensic Press test is a new product that detects saliva stains by reacting with amylase. When the paper is pressed against a saliva stain a blue spot occurs. To test the sensitivity of the paper, a set of dilution series of saliva down to 1:500 was prepared on cotton fabric. Blue spots could be seen for dilutions of

  16. Using the new Phadebas ® Forensic Press test to find crime scene saliva stains suitable for DNA analysis

    Microsoft Academic Search

    Johannes Hedman; Karin Gustavsson; Ricky Ansell

    2008-01-01

    The Phadebas® Forensic Press test is a new product that detects saliva stains by reacting with amylase. When the paper is pressed against a saliva stain a blue spot occurs. To test the sensitivity of the paper, a set of dilution series of saliva down to 1:500 was prepared on cotton fabric. Blue spots could be seen for dilutions of

  17. Screening of myocardial contraction bands: a comparison between two histological staining methods.

    PubMed

    Ståhl, E

    1990-03-01

    Two histological stains for demonstration of contraction bands (cb): Mallory's phosphotungstic acid hematoxylin (PTAH) and a modification of luxol fast blue (LFB) were compared. Microscopical sections from hearts from 60 autopsies were screened. The regional distribution of the cb lesions was also studied. It was found that PTAH showed cb more consistently than LFB. None of the stains were suitable for screening in the lowest magnification. In cases of cardiac death the cb were often extensive and scattered throughout the myocardium. In non-cardiac deaths with cb these were always discrete and seen only in one location which might be a means of differentiating lesions of pathogenetic importance from agonal ones. PMID:1692296

  18. A triple stain technique to evaluate monocyte, neutrophil, and eosinophil proliferation in soft agar cultures.

    PubMed

    Phillips, P G; Chikkappa, G; Brinson, P S

    1983-01-01

    A useful cytochemical technique has been developed which facilitates the identification of the types of cell aggregates which proliferate in soft agar cultures of hematopoietic stem cells. Staining of fixed agar discs for alpha-naphthyl acetate esterase (ANAE), naphthol AS-D chloroacetate esterase (CAE) and with Luxol fast blue (LFB) in sequence results in permanent preparations in which monocytic, neutrophilic and eosinophilic aggregates can readily be distinguished on the same slide. This represents an improvement over the currently used technique which relies on the removal of a representative sample of aggregates from the agar discs for cell typing. Whole plate staining results in more accurate estimates of eosinophil growth since the frequency of occurrence of these cell aggregates is low under many of the culturing conditions used. The technique provides a useful tool for studying normal and abnormal hematopoiesis. PMID:6187590

  19. Rates of Benthic Protozoan Grazing on Free and Attached Sediment Bacteria Measured with Fluorescently Stained Sediment

    PubMed Central

    Starink, Mathieu; Krylova, Irina N.; Bär-Gilissen, Marie-José; Bak, Rolf P. M.; Cappenberg, Thomas E.

    1994-01-01

    In order to determine the importance of benthic protozoa as consumers of bacteria, grazing rates have been measured by using monodispersed fluorescently labeled bacteria (FLB). However, high percentages of nongrazing benthic protists are reported in the literature. These are related to serious problems of the monodispersed FLB method. We describe a new method using 5-(4,6-dichlorotriazin-2-yl)-aminofluorescein (DTAF)-stained sediment to measure in situ bacterivory by benthic protists. This method is compared with the monodispersed FLB technique. Our estimates of benthic bacterivory range from 61 to 73 bacteria protist-1 h-1 and are about twofold higher than the results of the monodispersed FLB method. The number of nongrazing protists after incubation for 15 min with DTAF-stained sediment is in agreement with theoretical expectation. We also tested the relative affinity for FLB of protists and discuss the results with respect to a grazing model. PMID:16349315

  20. Visualization of latent blood stains using visible reflectance hyperspectral imaging and chemometrics.

    PubMed

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

    2015-01-01

    The detection of latent traces is an important aspect of crime scene investigation. Blood stains on black backgrounds can be visualized using chemiluminescence, which is invasive and requires a darkened room, or near-infrared photography, for which investigators need to change filters manually to optimize contrast. We demonstrated the performance of visible reflectance hyperspectral imaging (400-720 nm) for this purpose. Several processing methods were evaluated: single wavelength bands, ratio images, principal component analysis (PCA), and "SIMPLe-to-use Interactive Self-modeling Mixture Analysis" (SIMPLISMA). Using these methods, we were able to enhance the contrast between blood stains and 12 different fabrics. On black cotton, blood dilutions were visible with a minimal concentration of 25% of whole blood. The hyperspectral camera system used in this study is portable and wireless, which makes it suitable for crime scene use. The described technique is noncontact and nondestructive, so all traces are preserved for further analysis. PMID:25382735