Selenium Incorporated Cationic Organochalcogen: Live Cell Compatible and Highly Photostable Molecular Stain for Imaging and Localization of Intracellular DNA.

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Gaur, Pankaj; Kumar, Ajay; Dey, Gourab; Kumar, Rajendra; Bhattacharyya, Shalmoli; Ghosh, Subrata

2016-05-04

Successful integration of selenium unit into a newly designed cationic chemical architecture led to the development of a highly photostable molecular maker PA5 to be used in fluorescence microscopy as cellular nucleus staining agent for longer duration imaging under continuous laser illumination. Adaptation of a targeted single-atom modification strategy led to the development of a series of proficient DNA light-up probes (PA1-PA5). Further, their comparative photophysical studies in the presence of DNA revealed the potential of electron rich heteroatoms of chalcogen family in improving binding efficiency and specificity of molecular probes toward DNA. The findings of cell studies confirmed the outstanding cell compatibility of probe PA5 in terms of cell permeability, biostability, and extremely low cytotoxicity. Moreover, the photostability experiment employing continuous laser illumination in solution phase as well as in cell assay (both fixed and live cells) revealed the admirable photobleaching resistance of PA5. Finally, while investigating the phototoxicity of PA5, the probe was found not to exhibit light-induced toxicity even when irradiated for longer duration. All these experimental results demonstrated the promising standing of PA5 as a futuristic cell compatible potential stain for bioimaging and temporal profiling of DNA.

Highly increased detection of silver stained protein bands in polyacrylamide gels with thermo-optical methods

Science.gov (United States)

Mazza, Giulia; Posnicek, Thomas; Brandl, Martin

2016-11-01

Sodium dodecyl sulfate polyacrylamide gel electrophoresis is a well-known technique to separate proteins by their molecular weight. After electrophoresis, the gels are commonly stained for protein band analysis with silver stain; this allows the detection of protein loads to about 1 ng. To increase the detection sensitivity of the protein bands down in the subnanogram level, a sensor has been developed based on the thermal lens effect to scan and quantify protein loads which would remain undetected using the standard imaging systems. The thermal lens sensor is equipped with a 450 nm diode pump laser modulated at 1 Hz and a HeNe probe laser mounted in collinear geometry. The sensor could detect protein bands of 0.05 ng when the gel was soaked in methanol/water and 0.1 ng in water. The limit of detection ranged from 8 to 20 pg, depending on the soaking medium and the staining efficiency. Thus, the detection of silver stain by thermal lens effect results 10 to 20 times more sensitive than the standard colorimetric method.

Probing Zeolite Crystal Architecture and Structural Imperfections using Differently Sized Fluorescent Organic Probe Molecules.

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Hendriks, Frank C; Schmidt, Joel E; Rombouts, Jeroen A; Lammertsma, Koop; Bruijnincx, Pieter C A; Weckhuysen, Bert M

2017-05-05

A micro-spectroscopic method has been developed to probe the accessibility of zeolite crystals using a series of fluorescent 4-(4-diethylaminostyryl)-1-methylpyridinium iodide (DAMPI) probes of increasing molecular size. Staining large zeolite crystals with MFI (ZSM-5) topology and subsequent mapping of the resulting fluorescence using confocal fluorescence microscopy reveal differences in structural integrity: the 90° intergrowth sections of MFI crystals are prone to develop structural imperfections, which act as entrance routes for the probes into the zeolite crystal. Polarization-dependent measurements provide evidence for the probe molecule's alignment within the MFI zeolite pore system. The developed method was extended to BEA (Beta) crystals, showing that the previously observed hourglass pattern is a general feature of BEA crystals with this morphology. Furthermore, the probes can accurately identify at which crystal faces of BEA straight or sinusoidal pores open to the surface. The results show this method can spatially resolve the architecture-dependent internal pore structure of microporous materials, which is difficult to assess using other characterization techniques such as X-ray diffraction. © 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

Stain Deconvolution Using Statistical Analysis of Multi-Resolution Stain Colour Representation.

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Najah Alsubaie

Full Text Available Stain colour estimation is a prominent factor of the analysis pipeline in most of histology image processing algorithms. Providing a reliable and efficient stain colour deconvolution approach is fundamental for robust algorithm. In this paper, we propose a novel method for stain colour deconvolution of histology images. This approach statistically analyses the multi-resolutional representation of the image to separate the independent observations out of the correlated ones. We then estimate the stain mixing matrix using filtered uncorrelated data. We conducted an extensive set of experiments to compare the proposed method to the recent state of the art methods and demonstrate the robustness of this approach using three different datasets of scanned slides, prepared in different labs using different scanners.

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

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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

Double-staining chromogenic in situ hybridization as a useful alternative to split-signal fluorescence in situ hybridization in lymphoma diagnostics

DEFF Research Database (Denmark)

van Rijk, A.; Svenstroup-Poulsen, T.; Jones, M.

2010-01-01

within the reach of every pathology laboratory. Design and Methods Our study was initiated to determine the consistency between chromogenic in situ hybridization and fluorescence in situ hybridization, both using split-signal probes developed for the detection of chromosomal breaks. Five hundred...... and actual signal were compared to the original fluorescence hybridization results. In addition, hematoxylin background staining intensity and signal intensity of the double-staining chromogenic in situ hybridization procedure were analyzed. Results With respect to the presence or absence of chromosomal...

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

International Nuclear Information System (INIS)

Lee, Deok Hee; Hwang, Jae Cheol; Lim, Soo Mee; Yoon, Hyun-Ki; Sung, Kyu-Bo; Song, Ho-Young

2000-01-01

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

ATP-containing vesicles in stria vascular marginal cell cytoplasms in neonatal rat cochlea are lysosomes.

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Liu, Jun; Liu, Wenjing; Yang, Jun

2016-02-11

We confirmed that ATP is released from cochlear marginal cells in the stria vascular but the cell organelle in which ATP stores was not identified until now. Thus, we studied the ATP-containing cell organelles and suggest that these are lysosomes. Primary cultures of marginal cells of Sprague-Dawley rats aged 1-3 days was established. Vesicles within marginal cells stained with markers were identified under confocal laser scanning microscope and transmission electron microscope (TEM). Then ATP release from marginal cells was measured after glycyl-L-phenylalanine-ß- naphthylamide (GPN) treatment using a bioluminescent assay. Quinacrine-stained granules within marginal cells were labeled with LysoTracker, a lysosome tracer, and lysosomal-associated membrane protein 1(LAMP1), but not labeled with the mitochondrial tracer MitoTracker. Furthermore, LysoTracker-labelled puncta showed accumulation of Mant-ATP, an ATP analog. Treatment with 200 μM GPN quenched fluorescently labeled puncta after incubation with LysoTracker or quinacrine, but not MitoTracker. Quinacrine-labeled organelles observed by TEM were lysosomes, and an average 27.7 percent increase in ATP luminescence was observed in marginal cells extracellular fluid after GPN treatment. ATP-containing vesicles in cochlear marginal cells of the stria vascular from neonatal rats are likely lysosomes. ATP release from marginal cells may be via Ca(2+)-dependent lysosomal exocytosis.

Neural stem cells for disease modeling of Wolman disease and evaluation of therapeutics.

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Aguisanda, Francis; Yeh, Charles D; Chen, Catherine Z; Li, Rong; Beers, Jeanette; Zou, Jizhong; Thorne, Natasha; Zheng, Wei

2017-06-28

Wolman disease (WD) is a rare lysosomal storage disorder that is caused by mutations in the LIPA gene encoding lysosomal acid lipase (LAL). Deficiency in LAL function causes accumulation of cholesteryl esters and triglycerides in lysosomes. Fatality usually occurs within the first year of life. While an enzyme replacement therapy has recently become available, there is currently no small-molecule drug treatment for WD. We have generated induced pluripotent stem cells (iPSCs) from two WD patient dermal fibroblast lines and subsequently differentiated them into neural stem cells (NSCs). The WD NSCs exhibited the hallmark disease phenotypes of neutral lipid accumulation, severely deficient LAL activity, and increased LysoTracker dye staining. Enzyme replacement treatment dramatically reduced the WD phenotype in these cells. In addition, δ-tocopherol (DT) and hydroxypropyl-beta-cyclodextrin (HPBCD) significantly reduced lysosomal size in WD NSCs, and an enhanced effect was observed in DT/HPBCD combination therapy. The results demonstrate that these WD NSCs are valid cell-based disease models with characteristic disease phenotypes that can be used to evaluate drug efficacy and screen compounds. DT and HPBCD both reduce LysoTracker dye staining in WD cells. The cells may be used to further dissect the pathology of WD, evaluate compound efficacy, and serve as a platform for high-throughput drug screening to identify new compounds for therapeutic development.

Gram staining with an automatic machine.

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Felek, S; Arslan, A

1999-01-01

This study was undertaken to develop a new Gram-staining machine controlled by a micro-controller and to investigate the quality of slides that were stained in the machine. The machine was designed and produced by the authors. It uses standard 220 V AC. Staining, washing, and drying periods are controlled by a timer built in the micro-controller. A software was made that contains a certain algorithm and time intervals for the staining mode. One-hundred and forty smears were prepared from Escherichia coli, Staphylococcus aureus, Neisseria sp., blood culture, trypticase soy broth, direct pus and sputum smears for comparison studies. Half of the slides in each group were stained with the machine, the other half by hand and then examined by four different microbiologists. Machine-stained slides had a higher clarity and less debris than the hand-stained slides (p stained slides, some Gram-positive organisms showed poor Gram-positive staining features (p Gram staining with the automatic machine increases the staining quality and helps to decrease the work load in a busy diagnostic laboratory.

Bis-pyridinium quadrupolar derivatives. High Stokes shift selective probes for bio-imaging

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Salice, Patrizio; Versari, Silvia; Bradamante, Silvia; Meinardi, Francesco; Macchi, Giorgio; Pagani, Giorgio A.; Beverina, Luca

2013-11-01

We describe the design, synthesis and characterization of five high Stokes shift quadrupolar heteroaryl compounds suitable as fluorescent probes in bio-imaging. In particular, we characterize the photophysical properties and the intracellular localization in Human Umbilical Vein Endothelial Cells (HUVEC) and Human Mesenchymal Stem Cells (HMSCs) for each dye. We show that, amongst all of the investigated derivatives, the 2,5-bis[1-(4-N-methylpyridinium)ethen-2-yl)]- N-methylpyrrole salt is the best candidates as selective mitochondrial tracker. Finally, we recorded the full emission spectrum of the most performing - exclusively mitochondrial selective - fluorescent probe directly from HUVEC stained cells. The emission spectrum collected from the stained mitochondria shows a remarkably more pronounced vibronic structure with respect to the emission of the free fluorophore in solution.

A Thiazole Coumarin (TC) Turn-On Fluorescence Probe for AT-Base Pair Detection and Multipurpose Applications in Different Biological Systems

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Narayanaswamy, Nagarjun; Kumar, Manoj; Das, Sadhan; Sharma, Rahul; Samanta, Pralok K.; Pati, Swapan K.; Dhar, Suman K.; Kundu, Tapas K.; Govindaraju, T.

2014-01-01

Sequence-specific recognition of DNA by small turn-on fluorescence probes is a promising tool for bioimaging, bioanalytical and biomedical applications. Here, the authors report a novel cell-permeable and red fluorescent hemicyanine-based thiazole coumarin (TC) probe for DNA recognition, nuclear staining and cell cycle analysis. TC exhibited strong fluorescence enhancement in the presence of DNA containing AT-base pairs, but did not fluoresce with GC sequences, single-stranded DNA, RNA and proteins. The fluorescence staining of HeLa S3 and HEK 293 cells by TC followed by DNase and RNase digestion studies depicted the selective staining of DNA in the nucleus over the cytoplasmic region. Fluorescence-activated cell sorting (FACS) analysis by flow cytometry demonstrated the potential application of TC in cell cycle analysis in HEK 293 cells. Metaphase chromosome and malaria parasite DNA imaging studies further confirmed the in vivo diagnostic and therapeutic applications of probe TC. Probe TC may find multiple applications in fluorescence spectroscopy, diagnostics, bioimaging and molecular and cell biology. PMID:25252596

Accelerated staining technique using kitchen microwave oven

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Archana Mukunda

2015-01-01

Full Text Available Introduction: Histopathological diagnosis of specimens is greatly dependent on good sample preparation and staining. Both of these processes is governed by diffusion of fluids and dyes in and out of the tissue, which is the key to staining. Diffusion of fluids can be accelerated by the application of heat that reduces the time of staining from hours to the minute. We modified an inexpensive model of kitchen microwave oven for staining. This study is an attempt to compare the reliability of this modified technique against the tested technique of routine staining so as to establish the kitchen microwave oven as a valuable diagnostic tool. Materials and Methods: Sixty different tissue blocks were used to prepare 20 pairs of slides for 4 different stains namely hematoxylin and eosin, Van Gieson′s, 0.1% toluidine blue and periodic acid-Schiff. From each tissue block, two bits of tissues were mounted on two different slides. One slide was stained routinely, and the other stained inside a microwave. A pathologist evaluated the stained slides and the results so obtained were analyzed statistically. Results: Microwave staining considerably cut down the staining time from hours to seconds. Microwave staining showed no loss of cellular and nuclear details, uniform-staining characteristics and was of excellent quality. Interpretation and Conclusion: The cellular details, nuclear details and staining characteristics of microwave stained tissues were better than or equal to the routine stained tissue. The overall quality of microwave-stained sections was found to be better than the routine stained tissue in majority of cases.

Quantifying filamentous microorganisms in activated sludge before, during, and after an incident of foaming by oligonucleotide probe hybridizations and antibody staining.

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Oerther, D B; de los Reyes, F L; de los Reyes, M F; Raskin, L

2001-10-01

Quantitative oligonucleotide probe hybridizations, immunostaining, and a simple foaming potential test were used to follow an incident of seasonal filamentous foaming at the Urbana-Champaign Sanitary District, Northeast Wastewater Treatment Plant. A positive correlation was observed between an increase in foaming potential and the appearance of foam on the surfaces of aeration basins and secondary clarifiers. In addition, during the occurrence of foaming, the mass and activity of Gordonia spp. increased as measured by fluorescence in situ hybridization, antibody staining, and quantitative membrane hybridization of RNA extracts. An increase in Gordonia spp. rRNA levels from 0.25 to 1.4% of total rRNA was observed using quantitative membrane hybridizations, whereas during the same period, the fraction of mixed liquor volatile suspended solids attributed to Gordonia spp. increased from 4% to more than 32% of the total mixed liquor volatile suspended solids. These results indicate that both the activity and biomass level of Gordonia spp. in activated sludge increased relative to the activity aid the biomass level of the complete microbial community during a seasonal occurrence of filamentous foaming. Thus, Gordonia spp. may represent a numerically dominant but metabolically limited fraction of the total biomass, and the role of Gordonia spp. in filamentous foaming may be linked more tightly to the physical presence of filamentous microorganisms than to the metabolic activity of the cells.

Circulating Hemocytes from Larvae of the Japanese Rhinoceros Beetle Allomyrina dichotoma (Linnaeus) (Coleoptera: Scarabaeidae) and the Cellular Immune Response to Microorganisms.

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Hwang, Sejung; Bang, Kyeongrin; Lee, Jiae; Cho, Saeyoull

2015-01-01

Hemocytes of the last larva of the Japanese rhinoceros beetle A. dichotoma (Linnaeus) (Coleoptera: Scarabaeidae) were classified as granulocytes, plasmatocytes, oenocytoids, spherulocytes, prohemocytes, and adipohemocytes. Among these cell types, only the granulocytes became immunologically activated with obvious morphological changes, displaying large amoeba-like, lobopodia-like, and fan-like structures. In addition, their cytoplasmic granules became larger and greatly increased in number. To explore whether these granules could be immunologically generated as phagosomes, total hemocytes were stained with LysoTracker. Greater than 90% of the granulocytes retained the LysoTracker dye at 4 h post-bacterial infection. In flow cytometry analysis, the red fluorescent signal was highly increased at 4 h post-bacterial infection (60.36%) compared to controls (5.08%), as was confirmed by fluorescent microscopy. After 12 h post-infection, these signals returned to basal levels. The uptake of pathogens by granulocytes rapidly triggered the translocation of the microtubule-associated protein 1 light chain 3 alpha (LC3) to the phagosome, which may result in enhanced pathogen killing.

Circulating Hemocytes from Larvae of the Japanese Rhinoceros Beetle Allomyrina dichotoma (Linnaeus (Coleoptera: Scarabaeidae and the Cellular Immune Response to Microorganisms.

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Sejung Hwang

Full Text Available Hemocytes of the last larva of the Japanese rhinoceros beetle A. dichotoma (Linnaeus (Coleoptera: Scarabaeidae were classified as granulocytes, plasmatocytes, oenocytoids, spherulocytes, prohemocytes, and adipohemocytes. Among these cell types, only the granulocytes became immunologically activated with obvious morphological changes, displaying large amoeba-like, lobopodia-like, and fan-like structures. In addition, their cytoplasmic granules became larger and greatly increased in number. To explore whether these granules could be immunologically generated as phagosomes, total hemocytes were stained with LysoTracker. Greater than 90% of the granulocytes retained the LysoTracker dye at 4 h post-bacterial infection. In flow cytometry analysis, the red fluorescent signal was highly increased at 4 h post-bacterial infection (60.36% compared to controls (5.08%, as was confirmed by fluorescent microscopy. After 12 h post-infection, these signals returned to basal levels. The uptake of pathogens by granulocytes rapidly triggered the translocation of the microtubule-associated protein 1 light chain 3 alpha (LC3 to the phagosome, which may result in enhanced pathogen killing.

Investigation of the Application of miR10b and miR135b in the Identification of Semen Stains

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Xue, Tianyu; Ma, Xiaoyan; Zhang, Jinxiang; Ou, Xueling; Cheng, Jianding; Sun, Hongyu

2015-01-01

To evaluate the identification method using the microRNA markers miR10b and miR135b to distinguish semen stains from menstrual blood, peripheral blood, vaginal fluid and so on body fluid stains. The expression levels of miR10b and miR35b in semen stains and menstrual blood and so on were detected utilizing a real-time quantitative PCR technique with a specific fluorescence-labeled TaqMan probe. RNU6b was used as the internal reference gene; the difference in their expression was analyzed, and the specificity, sensitivity, and detection capability of the techniques were evaluated. The expression of miR10b and miR135b in semen stains was significantly higher than that of other body fluid stains, with a mean value of ΔCт from-6 to-7. However, it ranged from-2 to-4 for other body fluid stains. The initial criteria for judging which semen stains can be identified were determined by analyzing the research results. When the threshold value was set to 0.04, the CT value could be detected in the target genes miR10b, miR135b and in the internal reference gene RNU6b, and CT values are<40, ΔCT[10b-U6]<-5.5, and ΔCT[135b-U6]<-6, respectively, and the semen stain could be identified. The expression levels of miR10b and miR135b are higher in semen with strong tissue specificity; thus, they can be used to differentiate semen stains from other body fluid stains in forensic science. PMID:26355456

Investigation of the Application of miR10b and miR135b in the Identification of Semen Stains.

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Dayue Tong

Full Text Available To evaluate the identification method using the microRNA markers miR10b and miR135b to distinguish semen stains from menstrual blood, peripheral blood, vaginal fluid and so on body fluid stains. The expression levels of miR10b and miR35b in semen stains and menstrual blood and so on were detected utilizing a real-time quantitative PCR technique with a specific fluorescence-labeled TaqMan probe. RNU6b was used as the internal reference gene; the difference in their expression was analyzed, and the specificity, sensitivity, and detection capability of the techniques were evaluated. The expression of miR10b and miR135b in semen stains was significantly higher than that of other body fluid stains, with a mean value of ΔCт from-6 to-7. However, it ranged from-2 to-4 for other body fluid stains. The initial criteria for judging which semen stains can be identified were determined by analyzing the research results. When the threshold value was set to 0.04, the CT value could be detected in the target genes miR10b, miR135b and in the internal reference gene RNU6b, and CT values are<40, ΔCT[10b-U6]<-5.5, and ΔCT[135b-U6]<-6, respectively, and the semen stain could be identified. The expression levels of miR10b and miR135b are higher in semen with strong tissue specificity; thus, they can be used to differentiate semen stains from other body fluid stains in forensic science.

Heavy ion irradiation induces autophagy in irradiated C2C12 myoblasts and their bystander cells

International Nuclear Information System (INIS)

Hino, Mizuki; Tajika, Yuki; Hamada, Nobuyuki

2010-01-01

Autophagy is one of the major processes involved in the degradation of intracellular materials. Here, we examined the potential impact of heavy ion irradiation on the induction of autophagy in irradiated C2C12 mouse myoblasts and their non-targeted bystander cells. In irradiated cells, ultrastructural analysis revealed the accumulation of autophagic structures at various stages of autophagy (id est (i.e.) phagophores, autophagosomes and autolysosomes) within 20 min after irradiation. Multivesicular bodies (MVBs) and autolysosomes containing MVBs (amphisomes) were also observed. Heavy ion irradiation increased the staining of microtubule-associated protein 1 light chain 3 and LysoTracker Red (LTR). Such enhanced staining was suppressed by an autophagy inhibitor 3-methyladenine. In addition to irradiated cells, bystander cells were also positive with LTR staining. Altogether, these results suggest that heavy ion irradiation induces autophagy not only in irradiated myoblasts but also in their bystander cells. (author)

Detection of stain formation on teeth by oral antiseptic solution using fiber optic displacement sensor

Science.gov (United States)

Rahman, H. A.; Rahim, H. R. A.; Harun, S. W.; Yasin, M.; Apsari, R.; Ahmad, H.; Wan Abas, W. A. B.

2013-02-01

The application of a simple intensity modulated fiber optic displacement sensor for the detection of stain formation on human teeth is demonstrated. The proposed sensor uses a concentric type bundled plastic optical fiber (POF) as a probe in conjunction with the surfaces of five human teeth as the reflecting targets. Prior to the experiment, the stains were produced extrinsically by soaking the teeth in different concentrations of oral antiseptic solution containing hexetidine. The concentration of the oral antiseptic solution is measured in volume%. For a concentration change from 0% to 80%, the peak voltage decreases exponentially from 1.15 mV to 0.41 mV with a measured resolution of 0.48% and 1.75% for concentration ranges of 0-40% and 40-80%, respectively. The correlation between the detector output and variation in the color of human tooth surface has successfully been examined. Simple in design and low in cost, this sensor can detect color changes due to hexetidine-induced stain on a tooth surface in a fast and convenient way. Thus, this sensor will be very promising in esthetic dentistry, dental color matching techniques, chemical and biomedical applications.

Smart optical probes for near-infrared fluorescence imaging of Alzheimer's disease pathology

International Nuclear Information System (INIS)

Raymond, Scott B.; Bacskai, Brian J.; Skoch, Jesse; Hills, Ivory D.; Swager, Timothy M.; Nesterov, Evgueni E.

2008-01-01

Near-infrared fluorescent probes for amyloid-beta (Aβ) are an exciting option for molecular imaging in Alzheimer's disease research and may translate to clinical diagnostics. However, Aβ-targeted optical probes often suffer from poor specificity and slow clearance from the brain. We are designing smart optical probes that emit characteristic fluorescence signal only when bound to Aβ. We synthesized a family of dyes and tested Aβ-binding sensitivity with fluorescence spectroscopy and tissue-staining. Select compounds exhibited Aβ-dependent changes in fluorescence quantum yield, lifetime, and emission spectra that may be imaged microscopically or in vivo using new lifetime and spectral fluorescence imaging techniques. Smart optical probes that turn on when bound to Aβ will improve amyloid detection and may enable quantitative molecular imaging in vivo. (orig.)

A new fluorescent pH probe for imaging lysosomes in living cells.

Science.gov (United States)

Lv, Hong-Shui; Huang, Shu-Ya; Xu, Yu; Dai, Xi; Miao, Jun-Ying; Zhao, Bao-Xiang

2014-01-15

A new rhodamine B-based pH fluorescent probe has been synthesized and characterized. The probe responds to acidic pH with short response time, high selectivity and sensitivity, and exhibits a more than 20-fold increase in fluorescence intensity within the pH range of 7.5-4.1 with the pKa value of 5.72, which is valuable to study acidic organelles in living cells. Also, it has been successfully applied to HeLa cells, for its low cytotoxicity, brilliant photostability, good membrane permeability and no 'alkalizing effect' on lysosomes. The results demonstrate that this probe is a lysosome-specific probe, which can selectively stain lysosomes and monitor lysosomal pH changes in living cells. Copyright © 2013 Elsevier Ltd. All rights reserved.

Iron Stain on Wood

Science.gov (United States)

Mark Knaebe

2013-01-01

Iron stain, an unsightly blue–black or gray discoloration, can occur on nearly all woods. Oak, redwood, cypress, and cedar are particularly prone to iron stain because these woods contain large amounts of tannin-like extractives. The discoloration is caused by a chemical reaction between extractives in the wood and iron in steel products, such as nails, screws, and...

Demonstration of lipofuscin and Nissl bodies in crystal violet stained sections using a fluorescence technique or pyronin Y stain.

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Terr, L I

1986-09-01

This paper presents two simple, reliable methods for identification of lipofuscin and Nissl bodies in the same section. One method shows that lipofuscin stained with crystal violet retains its ability to fluoresce and can be observed under the fluorescence microscope after the stain has faded. Fading is accompanied by a gradual increase in the intensity of the fluorescence and is complete in about 5 min. Exciting illumination from this part of the spectrum also substantially fades staining of other autofluorescing tissue elements, such as lipids. Nonfluorescing structures, such as Nissl bodies, remain stained. By changing from transillumination with tungsten light to epifluorescent illumination and vice versa, both types of structures--Nissl bodies and lipofuscin--can be identified in the same section. The second technique uses pyronin Y for staining Nissl bodies in preparations previously stained with crystal violet. Nissl bodies are stained pink but lipofuscin remains violet. Lipofuscin in these sections also remains autofluorescent after the crystal violet stain has faded under violet or near-UV light.

A novel staining protocol for multiparameter assessment of cell heterogeneity in Phormidium populations (cyanobacteria employing fluorescent dyes.

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Daria Tashyreva

Full Text Available Bacterial populations display high heterogeneity in viability and physiological activity at the single-cell level, especially under stressful conditions. We demonstrate a novel staining protocol for multiparameter assessment of individual cells in physiologically heterogeneous populations of cyanobacteria. The protocol employs fluorescent probes, i.e., redox dye 5-cyano-2,3-ditolyl tetrazolium chloride, 'dead cell' nucleic acid stain SYTOX Green, and DNA-specific fluorochrome 4',6-diamidino-2-phenylindole, combined with microscopy image analysis. Our method allows simultaneous estimates of cellular respiration activity, membrane and nucleoid integrity, and allows the detection of photosynthetic pigments fluorescence along with morphological observations. The staining protocol has been adjusted for, both, laboratory and natural populations of the genus Phormidium (Oscillatoriales, and tested on 4 field-collected samples and 12 laboratory strains of cyanobacteria. Based on the mentioned cellular functions we suggest classification of cells in cyanobacterial populations into four categories: (i active and intact; (ii injured but active; (iii metabolically inactive but intact; (iv inactive and injured, or dead.

Histological Stains: A Literature Review and Case Study.

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Alturkistani, Hani A; Tashkandi, Faris M; Mohammedsaleh, Zuhair M

2015-06-25

The history of histology indicates that there have been significant changes in the techniques used for histological staining through chemical, molecular biology assays and immunological techniques, collectively referred to as histochemistry. Early histologists used the readily available chemicals to prepare tissues for microscopic studies; these laboratory chemicals were potassium dichromate, alcohol and the mercuric chloride to harden cellular tissues. Staining techniques used were carmine, silver nitrate, Giemsa, Trichrome Stains, Gram Stain and Hematoxylin among others. The purpose of this research was to assess past and current literature reviews, as well as case studies, with the aim of informing ways in which histological stains have been improved in the modern age. Results from the literature review has indicated that there has been an improvement in histopathology and histotechnology in stains used. There has been a rising need for efficient, accurate and less complex staining procedures. Many stain procedures are still in use today, and many others have been replaced with new immunostaining, molecular, non-culture and other advanced staining techniques. Some staining methods have been abandoned because the chemicals required have been medically proven to be toxic. The case studies indicated that in modern histology a combination of different stain techniques are used to enhance the effectiveness of the staining process. Currently, improved histological stains, have been modified and combined with other stains to improve their effectiveness.

Two rhodamine lactam modulated lysosome-targetable fluorescence probes for sensitively and selectively monitoring subcellular organelle pH change

Energy Technology Data Exchange (ETDEWEB)

Li, Hongmei [Ministry of Education Key Laboratory of Synthetic and Natural Functional Molecule Chemistry, College of Chemistry & Materials Science, Northwest University, Xi' an 710069 (China); Wang, Cuiling [Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi' an 710069 (China); She, Mengyao; Zhu, Yuelu; Zhang, Jidong; Yang, Zheng [Ministry of Education Key Laboratory of Synthetic and Natural Functional Molecule Chemistry, College of Chemistry & Materials Science, Northwest University, Xi' an 710069 (China); Liu, Ping, E-mail: liuping@nwu.edu.cn [Ministry of Education Key Laboratory of Synthetic and Natural Functional Molecule Chemistry, College of Chemistry & Materials Science, Northwest University, Xi' an 710069 (China); Wang, Yaoyu [Ministry of Education Key Laboratory of Synthetic and Natural Functional Molecule Chemistry, College of Chemistry & Materials Science, Northwest University, Xi' an 710069 (China); Li, Jianli, E-mail: lijianli@nwu.edu.cn [Ministry of Education Key Laboratory of Synthetic and Natural Functional Molecule Chemistry, College of Chemistry & Materials Science, Northwest University, Xi' an 710069 (China)

2015-11-05

Be a powerful technique for convenient detection of pH change in living cells, especially at subcellular level, fluorescent probes has attracted more and more attention. In this work, we designed and synthesized three rhodamine lactam modulated fluorescent probes RS1, RS2 and RS3, which all respond sensitively toward weak acidity (pH range 4–6) via the photophysical property in buffer solution without interference from the other metal ions, and they also show ideal pKa values and excellent reversibility. Particularly, by changing the lone pair electrons distribution of lactam-N atom with different conjugations, RS2 and RS3 exhibit high quantum yield, negligible cytotoxicity and excellent permeability. They are suitable to stain selectively lysosomes of tumor cells and monitor its pH changes sensitively via optical molecular imaging. The above findings suggest that the probes we designed could act as ideal and easy method for investigating the pivotal role of H{sup +} in lysosomes and are potential pH detectors in disease diagnosis through direct intracellular imaging. - Highlights: • Two probes for sensitively and selectively monitoring weak acidic pH change. • The pKa of the probes was highly suitable for staining lysosomes in tumor cells. • The properties of those probes were changed by different conjugate system. • These probes have negligible cytotoxicity and good sensitivity in vivo.

Two rhodamine lactam modulated lysosome-targetable fluorescence probes for sensitively and selectively monitoring subcellular organelle pH change

International Nuclear Information System (INIS)

Li, Hongmei; Wang, Cuiling; She, Mengyao; Zhu, Yuelu; Zhang, Jidong; Yang, Zheng; Liu, Ping; Wang, Yaoyu; Li, Jianli

2015-01-01

Be a powerful technique for convenient detection of pH change in living cells, especially at subcellular level, fluorescent probes has attracted more and more attention. In this work, we designed and synthesized three rhodamine lactam modulated fluorescent probes RS1, RS2 and RS3, which all respond sensitively toward weak acidity (pH range 4–6) via the photophysical property in buffer solution without interference from the other metal ions, and they also show ideal pKa values and excellent reversibility. Particularly, by changing the lone pair electrons distribution of lactam-N atom with different conjugations, RS2 and RS3 exhibit high quantum yield, negligible cytotoxicity and excellent permeability. They are suitable to stain selectively lysosomes of tumor cells and monitor its pH changes sensitively via optical molecular imaging. The above findings suggest that the probes we designed could act as ideal and easy method for investigating the pivotal role of H + in lysosomes and are potential pH detectors in disease diagnosis through direct intracellular imaging. - Highlights: • Two probes for sensitively and selectively monitoring weak acidic pH change. • The pKa of the probes was highly suitable for staining lysosomes in tumor cells. • The properties of those probes were changed by different conjugate system. • These probes have negligible cytotoxicity and good sensitivity in vivo.

Molecular Imaging of β-Amyloid Plaques with Near-Infrared Boron Dipyrromethane (BODIPY-Based Fluorescent Probes

Directory of Open Access Journals (Sweden)

Hiroyuki Watanabe

2013-07-01

Full Text Available The formation of β-amyloid (Aβ plaques is a critical neurodegenerative change in Alzheimer disease (AD. We designed and synthesized novel boron dipyrromethane (BODIPY-based Aβ probes (BAPs and evaluated their utility for near-infrared fluorescence imaging of Aβ plaques in the brain. In binding experiments in vitro, BAPs showed high affinity for synthetic Aβ aggregates (Kd = 18–149 nM. Furthermore, BAPs clearly stained Aβ plaques in sections of Tg2576 mice. In mouse brain tissue, BAPs showed sufficient uptake for optical imaging. In addition, ex vivo fluorescent staining of brain sections from Tg2576 mice after the injection of BAP-2 showed selective binding of Aβ plaques with little nonspecific binding. BAPs may be useful as a near-infrared fluorescent probe for imaging Aβ plaques.

Centrifuge-operated specimen staining method and apparatus

Science.gov (United States)

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.

Selection and application of exterior stains for wood

Science.gov (United States)

R. Sam. Williams; William C. Feist

1999-01-01

Exterior stains for wood protect the wood surface from sunlight and moisture. Because stains are formulated to penetrate the wood surface, they are not prone to crack or peel as can film-forming finishes, such as paints. This publication describes the properties of stains and wood, methods for applying stains, and the expected service life of stains.

Differential staining of bacteria: acid fast stain.

Science.gov (United States)

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

2009-11-01

Acid-fastness is an uncommon characteristic shared by the genera Mycobacterium (Section 10A) and Nocardia. Because of this feature, this stain is extremely helpful in identification of these bacteria. Although Gram positive, acid-fast bacteria do not take the crystal violet into the wall well, appearing very light purple rather than the deep purple of normal Gram-positive bacteria. (c) 2009 by John Wiley & Sons, Inc.

FLOW CYTOMETRIC APPLICABILITY OF FLUORESCENT VITALITY PROBES ON PHYTOPLANKTON1.

Science.gov (United States)

Peperzak, Louis; Brussaard, Corina P D

2011-06-01

The applicability of six fluorescent probes (four esterase probes: acetoxymethyl ester of Calcein [Calcein-AM], 5-chloromethylfluorescein diacetate [CMFDA], fluorescein diacetate [FDA], and 2',7'-dichlorofluorescein diacetate [H 2 DCFDA]; and two membrane probes: bis-(1,3-dibutylbarbituric acid) trimethine oxonol [DiBAC 4 (3)] and SYTOX-Green) as vitality stains was tested on live and killed cells of 40 phytoplankton strains in exponential and stationary growth phases, belonging to 12 classes and consisting of four cold-water, 26 temperate, and four warm-water species. The combined live/dead ratios of all six probes indicated significant differences between the 12 plankton classes (P live/dead ratios of FDA and CMFDA were not significantly different from each other, and both performed better than Calcein-AM and H 2 DCFDA (P live/dead ratios) among all six probes belonged to nine genera from six classes of phytoplankton. In conclusion, FDA, CMFDA, DIBAC 4 (3), and SYTOX-Green represent a wide choice of vitality probes in the study of phytoplankton ecology, applicable in many species from different algal classes, originating from different regions and at different stages of growth. © 2011 Phycological Society of America.

The comparison of pyrosequencing molecular Gram stain, culture, and conventional Gram stain for diagnosing orthopaedic infections.

Science.gov (United States)

Kobayashi, Naomi; Bauer, Thomas W; Tuohy, Marion J; Lieberman, Isador H; Krebs, Viktor; Togawa, Daisuke; Fujishiro, Takaaki; Procop, Gary W

2006-08-01

We have developed a combined real-time PCR and pyrosequencing assay that successfully differentiated the vast majority of gram-positive and gram-negative bacteria when bacterial isolates were tested. The purpose of this study was to evaluate this assay on clinical specimens obtained from orthopedic surgeries, and to prospectively compare the results of "molecular Gram stain" with culture and conventional direct Gram stain. Forty-five surgical specimens were obtained from patients who underwent orthopedic surgery procedures. The DNA was extracted and a set of broad-range PCR primers that targeted a part of the 16S rDNA gene was used for pan-bacterial PCR. The amplicons were submitted for pyrosequencing and the resulting molecular Gram stain characteristics were recorded. Culture and direct Gram staining were performed using standard methods for all cases. Surgical specimens were reviewed histologically for all cases that had a discrepancy between culture and molecular results. There was an 86.7% (39/45) agreement between the traditional and molecular methods. In 12/14 (85.7%) culture-proven cases of bacterial infection, molecular Gram stain characteristics were in agreement with the culture results, while the conventional Gram stain result was in agreement only for five cases (35.7%). In the 31 culture negative cases, 27 cases were also PCR negative, whereas 4 were PCR positive. Three of these were characterized as gram negative and one as gram positive by this molecular method. Molecular determination of the Gram stain characteristics of bacteria that cause orthopedic infections may be achieved, in most instances, by this method. Further studies are necessary to understand the clinical importance of PCR-positive/culture-negative results.

Metabolic flux ratio analysis and cell staining suggest the existence of C4 photosynthesis in Phaeodactylum tricornutum.

Science.gov (United States)

Huang, A; Liu, L; Zhao, P; Yang, C; Wang, G C

2016-03-01

Mechanisms for carbon fixation via photosynthesis in the diatom Phaeodactylum tricornutum Bohlin were studied recently but there remains a long-standing debate concerning the occurrence of C4 photosynthesis in this species. A thorough investigation of carbon metabolism and the evidence for C4 photosynthesis based on organelle partitioning was needed. In this study, we identified the flux ratios between C3 and C4 compounds in P. tricornutum using (13)C-labelling metabolic flux ratio analysis, and stained cells with various cell-permeant fluorescent probes to investigate the likely organelle partitioning required for single-cell C4 photosynthesis. Metabolic flux ratio analysis indicated the C3/C4 exchange ratios were high. Cell staining indicated organelle partitioning required for single-cell C4 photosynthesis might exist in P. tricornutum. The results of (13)C-labelling metabolic flux ratio analysis and cell staining suggest single-cell C4 photosynthesis exists in P. tricornutum. This study provides insights into photosynthesis patterns of P. tricornutum and the evidence for C4 photosynthesis based on (13)C-labelling metabolic flux ratio analysis and organelle partitioning. © 2015 The Society for Applied Microbiology.

Say goodbye to coffee stains

NARCIS (Netherlands)

Eral, Burak; van den Ende, Henricus T.M.; Mugele, Friedrich Gunther

2012-01-01

Discussing ideas over a mug of coffee or tea is the lifeblood of science, but have you ever thought about the stains that can be inadvertently left behind? H Burak Eral, Dirk van den Ende and Frieder Mugele explain how these stains, which can be a major annoyance in some biology techniques, can be

Diagnosing periprosthetic infection: false-positive intraoperative Gram stains.

Science.gov (United States)

Oethinger, Margret; Warner, Debra K; Schindler, Susan A; Kobayashi, Hideo; Bauer, Thomas W

2011-04-01

Intraoperative Gram stains have a reported low sensitivity but high specificity when used to help diagnose periprosthetic infections. In early 2008, we recognized an unexpectedly high frequency of apparent false-positive Gram stains from revision arthroplasties. The purpose of this report is to describe the cause of these false-positive test results. We calculated the sensitivity and specificity of all intraoperative Gram stains submitted from revision arthroplasty cases during a 3-month interval using microbiologic cultures of the same samples as the gold standard. Methods of specimen harvesting, handling, transport, distribution, specimen processing including tissue grinding/macerating, Gram staining, and interpretation were studied. After a test modification, results of specimens were prospectively collected for a second 3-month interval, and the sensitivity and specificity of intraoperative Gram stains were calculated. The retrospective review of 269 Gram stains submitted from revision arthroplasties indicated historic sensitivity and specificity values of 23% and 92%, respectively. Systematic analysis of all steps of the procedure identified Gram-stained but nonviable bacteria in commercial broth reagents used as diluents for maceration of periprosthetic membranes before Gram staining and culture. Polymerase chain reaction and sequencing showed mixed bacterial DNA. Evaluation of 390 specimens after initiating standardized Millipore filtering of diluent fluid revealed a reduced number of positive Gram stains, yielding 9% sensitivity and 99% specificity. Clusters of false-positive Gram stains have been reported in other clinical conditions. They are apparently rare related to diagnosing periprosthetic infections but have severe consequences if used to guide treatment. Even occasional false-positive Gram stains should prompt review of laboratory methods. Our observations implicate dead bacteria in microbiologic reagents as potential sources of false-positive Gram

Histopathological evaluation of ocular microsporidiosis by different stains

Directory of Open Access Journals (Sweden)

Sharma Savitri

2006-06-01

Full Text Available Abstract Background There is limited data on comparing stains in the detection of microsporidia in corneal biopsies. Hence we wanted to evaluate various stains for their ability to detect microsporidia in corneal tissue sections. Methods Four cases diagnosed with microsporidiosis on Hematoxylin and Eosin and Periodic Acid Schiff's stained sections of the corneal button between January 2002 and December 2004, were included. Further sections were prospectively stained with calcofluor white, Gram, Giemsa, Masson's trichrome, acridine orange, Gomori's methenamine silver, Gram's chromotrope and modified acid fast stain. The stained sections were analyzed for the spore characteristics in terms of size, shape, color contrast, cell wall morphology, waist band in cytoplasm and ease of detection. Results All sections showed microsporidial spores as 3 – 5 μm, oval bodies. 1% acid fast, Gram's chromotrope and GMS stains provided a reliable diagnosis of microsporidia as diagnostic waist band could be identified and good contrast helped distinguish the spores from inflammatory debris. Conclusion Considering the ease of performance, cost effectiveness and rapidity of the technique, 1% acid fast stain and Gram's chromotrope stain are ideal for the detection of microsporidia.

Validation of a simple and fast method to quantify in vitro mineralization with fluorescent probes used in molecular imaging of bone

International Nuclear Information System (INIS)

Moester, Martiene J.C.; Schoeman, Monique A.E.; Oudshoorn, Ineke B.; Beusekom, Mara M. van; Mol, Isabel M.; Kaijzel, Eric L.; Löwik, Clemens W.G.M.; Rooij, Karien E. de

2014-01-01

Highlights: •We validate a simple and fast method of quantification of in vitro mineralization. •Fluorescently labeled agents can detect calcium deposits in the mineralized matrix of cell cultures. •Fluorescent signals of the probes correlated with Alizarin Red S staining. -- Abstract: Alizarin Red S staining is the standard method to indicate and quantify matrix mineralization during differentiation of osteoblast cultures. KS483 cells are multipotent mouse mesenchymal progenitor cells that can differentiate into chondrocytes, adipocytes and osteoblasts and are a well-characterized model for the study of bone formation. Matrix mineralization is the last step of differentiation of bone cells and is therefore a very important outcome measure in bone research. Fluorescently labelled calcium chelating agents, e.g. BoneTag and OsteoSense, are currently used for in vivo imaging of bone. The aim of the present study was to validate these probes for fast and simple detection and quantification of in vitro matrix mineralization by KS483 cells and thus enabling high-throughput screening experiments. KS483 cells were cultured under osteogenic conditions in the presence of compounds that either stimulate or inhibit osteoblast differentiation and thereby matrix mineralization. After 21 days of differentiation, fluorescence of stained cultures was quantified with a near-infrared imager and compared to Alizarin Red S quantification. Fluorescence of both probes closely correlated to Alizarin Red S staining in both inhibiting and stimulating conditions. In addition, both compounds displayed specificity for mineralized nodules. We therefore conclude that this method of quantification of bone mineralization using fluorescent compounds is a good alternative for the Alizarin Red S staining

Validation of a simple and fast method to quantify in vitro mineralization with fluorescent probes used in molecular imaging of bone

Energy Technology Data Exchange (ETDEWEB)

Moester, Martiene J.C. [Department of Radiology, Leiden University Medical Center (Netherlands); Schoeman, Monique A.E. [Department of Orthopedic Surgery, Leiden University Medical Center (Netherlands); Oudshoorn, Ineke B. [Department of Radiology, Leiden University Medical Center (Netherlands); Percuros BV, Leiden (Netherlands); Beusekom, Mara M. van [Department of Radiology, Leiden University Medical Center (Netherlands); Mol, Isabel M. [Department of Radiology, Leiden University Medical Center (Netherlands); Percuros BV, Leiden (Netherlands); Kaijzel, Eric L.; Löwik, Clemens W.G.M. [Department of Radiology, Leiden University Medical Center (Netherlands); Rooij, Karien E. de, E-mail: k.e.de_rooij@lumc.nl [Department of Radiology, Leiden University Medical Center (Netherlands); Percuros BV, Leiden (Netherlands)

2014-01-03

Highlights: •We validate a simple and fast method of quantification of in vitro mineralization. •Fluorescently labeled agents can detect calcium deposits in the mineralized matrix of cell cultures. •Fluorescent signals of the probes correlated with Alizarin Red S staining. -- Abstract: Alizarin Red S staining is the standard method to indicate and quantify matrix mineralization during differentiation of osteoblast cultures. KS483 cells are multipotent mouse mesenchymal progenitor cells that can differentiate into chondrocytes, adipocytes and osteoblasts and are a well-characterized model for the study of bone formation. Matrix mineralization is the last step of differentiation of bone cells and is therefore a very important outcome measure in bone research. Fluorescently labelled calcium chelating agents, e.g. BoneTag and OsteoSense, are currently used for in vivo imaging of bone. The aim of the present study was to validate these probes for fast and simple detection and quantification of in vitro matrix mineralization by KS483 cells and thus enabling high-throughput screening experiments. KS483 cells were cultured under osteogenic conditions in the presence of compounds that either stimulate or inhibit osteoblast differentiation and thereby matrix mineralization. After 21 days of differentiation, fluorescence of stained cultures was quantified with a near-infrared imager and compared to Alizarin Red S quantification. Fluorescence of both probes closely correlated to Alizarin Red S staining in both inhibiting and stimulating conditions. In addition, both compounds displayed specificity for mineralized nodules. We therefore conclude that this method of quantification of bone mineralization using fluorescent compounds is a good alternative for the Alizarin Red S staining.

Conjugates of a Photoactivated Rhodamine with Biopolymers for Cell Staining

Science.gov (United States)

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

2014-01-01

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

Multicenter Assessment of Gram Stain Error Rates.

Science.gov (United States)

Samuel, Linoj P; Balada-Llasat, Joan-Miquel; Harrington, Amanda; Cavagnolo, Robert

2016-06-01

Gram stains remain the cornerstone of diagnostic testing in the microbiology laboratory for the guidance of empirical treatment prior to availability of culture results. Incorrectly interpreted Gram stains may adversely impact patient care, and yet there are no comprehensive studies that have evaluated the reliability of the technique and there are no established standards for performance. In this study, clinical microbiology laboratories at four major tertiary medical care centers evaluated Gram stain error rates across all nonblood specimen types by using standardized criteria. The study focused on several factors that primarily contribute to errors in the process, including poor specimen quality, smear preparation, and interpretation of the smears. The number of specimens during the evaluation period ranged from 976 to 1,864 specimens per site, and there were a total of 6,115 specimens. Gram stain results were discrepant from culture for 5% of all specimens. Fifty-eight percent of discrepant results were specimens with no organisms reported on Gram stain but significant growth on culture, while 42% of discrepant results had reported organisms on Gram stain that were not recovered in culture. Upon review of available slides, 24% (63/263) of discrepant results were due to reader error, which varied significantly based on site (9% to 45%). The Gram stain error rate also varied between sites, ranging from 0.4% to 2.7%. The data demonstrate a significant variability between laboratories in Gram stain performance and affirm the need for ongoing quality assessment by laboratories. Standardized monitoring of Gram stains is an essential quality control tool for laboratories and is necessary for the establishment of a quality benchmark across laboratories. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Research on pre-staining gel electrophoresis

International Nuclear Information System (INIS)

Zhong Ruibo; Liu Yushuang; Zhang Ping; Liu Jingran; Zhao Guofen; Zhang Feng

2014-01-01

Background: Gel electrophoresis is a powerful biochemical separation technique. Most biological molecules are completely transparent in the visible region of light, so it is necessary to use staining to show the results after gel electrophoresis, and the general steps of conventional staining methods are time-consuming. Purpose: We try to develop a novel approach to simplify the gel electrophoresis: Pre-Staining Gel Electrophoresis (PSGE), which can make the gel electrophoresis results monitored in real time. Methods: Pre-stain the protein samples with Coomassie Brilliant Blue (CBB) for 30 min before loading the sample into the gel well. Results and Conclusion: PSGE can be successfully used to analyze the binding efficiency of Bovine Serum Albumin (BSA) and amphiphilic polymer via chemical coupling and physical absorption, and the double PSGE also shows a great potential in bio-analytical chemistry. (authors)

Surface staining of small intestinal biopsies

DEFF Research Database (Denmark)

Poulsen, Steen Seier

1977-01-01

Small intestinal biopsies are most often by routine examined under a stereo-microscope, prior to embedding for histological examination. This is done in order to get a view of the appearance of the mucosal pattern, especially villus configuration. The distinctness of the surface pattern however......, is improved considerably if the biopsies are stained with Alcian Green and/or PAS before they are examined. In the present paper a detailed description is given of staining of small intestinal biopsies as whole mounts. The difference between the unstained and the stained biopsies is illustrated by a few...

Utility of Modified Ultrafast Papanicolaou Stain in Cytological Diagnosis.

Science.gov (United States)

Sinkar, Prachi; Arakeri, Surekha Ulhas

2017-03-01

Need for minimal turnaround time for assessing Fine Needle Aspiration Cytology (FNAC) has encouraged innovations in staining techniques that require lesser staining time with unequivocal cell morphology. The standard protocol for conventional Papanicolaou (PAP) stain requires about 40 minutes. To overcome this, Ultrafast Papanicolaou (UFP) stain was introduced which reduces staining time to 90 seconds and also enhances the quality. However, reagents required for this were not easily available hence, Modified Ultrafast Papanicolaou (MUFP) stain was introduced subsequently. To assess the efficacy of MUFP staining by comparing the quality of MUFP stain with conventional PAP stain. FNAC procedure was performed by using 10 ml disposable syringe and 22-23 G needle. Total 131 FNAC cases were studied which were lymph node (30), thyroid (38), breast (22), skin and soft tissue (24), salivary gland (11) and visceral organs (6). Two smears were prepared and stained by MUFP and conventional PAP stain. Scores were given on four parameters: background of smears, overall staining pattern, cell morphology and nuclear staining. Quality Index (QI) was calculated from ratio of total score achieved to maximum score possible. Statistical analysis using chi square test was applied to each of the four parameters before obtaining the QI in both stains. Students t-test was applied to evaluate the efficacy of MUFP in comparison with conventional PAP stain. The QI of MUFP for thyroid, breast, lymph node, skin and soft tissue, salivary gland and visceral organs was 0.89, 0.85, 0.89, 0.83, 0.92, and 0.78 respectively. Compared to conventional PAP stain QI of MUFP smears was better in all except visceral organ cases and was statistically significant. MUFP showed clear red blood cell background, transparent cytoplasm and crisp nuclear features. MUFP is fast, reliable and can be done with locally available reagents with unequivocal morphology which is the need of the hour for a cytopathology set-up.

Blood culture gram stain, acridine orange stain and direct sensitivity-based antimicrobial therapy of bloodstream infection in patients with trauma.

Science.gov (United States)

Behera, B; Mathur, P; Gupta, B

2010-01-01

The purpose of this study was to ascertain if the simple practice of Gram stain, acridine orange stain and direct sensitivity determination of positive blood culture bottles could be used to guide early and appropriate treatment in trauma patients with clinical suspicion of sepsis. The study also aimed to evaluate the error in interpreting antimicrobial sensitivity by direct method when compared to standard method and find out if specific antibiotic-organism combination had more discrepancies. Findings from consecutive episodes of blood stream infection at an Apex Trauma centre over a 12-month period are summarized. A total of 509 consecutive positive blood cultures were subjected to Gram staining. AO staining was done in BacT/ALERT-positive Gram-stain negative blood cultures. Direct sensitivity was performed from 369 blood culture broths, showing single type of growth in Gram and acridine orange staining. Results of direct sensitivity were compared to conventional sensitivity for errors. No 'very major' discrepancy was found in this study. About 5.2 and 1.8% minor error rates were noted in gram-positive and gram-negative bacteria, respectively, while comparing the two methods. Most of the discrepancies in gram-negative bacteria were noted in beta lactam - beta lactamase inhibitor combinations. Direct sensitivity testing was not reliable for reporting of methicillin and vancomycin resistance in Staphylococci. Gram stain result together with direct sensitivity testing is required for optimizing initial antimicrobial therapy in trauma patients with clinical suspicion of sepsis. Gram staining and AO staining proved particularly helpful in the early detection of candidaemia.

A nontoxic, photostable and high signal-to-noise ratio mitochondrial probe with mitochondrial membrane potential and viscosity detectivity

Science.gov (United States)

Chen, Yanan; Qi, Jianguo; Huang, Jing; Zhou, Xiaomin; Niu, Linqiang; Yan, Zhijie; Wang, Jianhong

2018-01-01

Herein, we reported a yellow emission probe 1-methyl-4-(6-morpholino-1, 3-dioxo-1H-benzo[de]isoquinolin-2(3H)-yl) pyridin-1-ium iodide which could specifically stain mitochondria in living immortalized and normal cells. In comparison to the common mitochondria tracker (Mitotracker Deep Red, MTDR), this probe was nontoxic, photostable and ultrahigh signal-to-noise ratio, which could real-time monitor mitochondria for a long time. Moreover, this probe also showed high sensitivity towards mitochondrial membrane potential and intramitochondrial viscosity change. Consequently, this probe was used for imaging mitochondria, detecting changes in mitochondrial membrane potential and intramitochondrial viscosity in physiological and pathological processes.

New Grocott Stain without Using Chromic Acid

International Nuclear Information System (INIS)

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

Gram staining apparatus for space station applications

Science.gov (United States)

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

1990-01-01

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

Efficacy of in-house fluorescent stain for fungus

Directory of Open Access Journals (Sweden)

K. R. L. Surya Kirani

2017-01-01

Full Text Available Context: Mycotic infections are gaining importance in the present day medicine, and definite demonstration of fungus is essential for diagnosis. Small numbers of organisms in the smear can be identified by fluorescence microscopy. Calcofluor white (CFW fluorescent stain is a textile brightener mixed with Evans blue. It is expensive and not easily available. Aims: (1 To assess the efficacy of in-house CFW fluorescent stain for fungus in relation to conventional CFW stain, histopathology, and culture. (2 To determine sensitivity, specificity, negative predictive value (NPV, and positive predictive value (PPV with culture as gold standard. Settings and Design: One hundred cases of suspected dermatophytosis and 15 cases of systemic mycosis were included in the study. Subjects and Methods: The local whitener Ranipal is added with Robin blue, another brightener, and was used to stain teased fungal cultures. Skin, hair, and nails require pretreatment with potassium hydroxide (KOH. Biopsy slides require deparaffinization and pretreatment with KOH before staining. Conventional calcofluor stain, histopathology, and culture were done. Statistical Analysis Used: Statistical analysis was performed using sensitivity, specificity, NPV, and PPV. Results: The results are consistently comparable with conventional stain. The sensitivity was 100%, specificity was 93.3%, NPV was 100%, and PPV was 85.7%. It is also cost effective when compared to commercial stains. Conclusions: In-house stain can be used for screening of fungus in direct samples, biopsies as alternative in resource-constrained laboratories.

Modified Genta triple stain for identifying Helicobacter pylori.

OpenAIRE

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 interf...

Comparison of staining of mitotic figures by haematoxylin and eosin-and crystal violet stains, in oral epithelial dysplasia and squamous cell carcinoma

Directory of Open Access Journals (Sweden)

Ankle Madhuri

2007-01-01

Full Text Available Mitosis of cells gives rise to tissue integrity. Defects during mitosis bring about abnormalities. Excessive proliferation of cells due to increased mitosis is one such outcome, which is the hallmark in precancer and cancer. The localization of proliferating cells or their precursors may not be obvious and easy. Establishing an easy way to distinguish these mitotic cells will help in grading and understanding their biological potential. Although immunohistochemistry is an advanced method in use, the cost and time factor makes it less feasible for many laboratories. Selective histochemical stains like toluidine blue, giemsa and crystal violet have been used in tissues including the developing brain, neural tissue and skin. Aim of the study: 1To compare the staining of mitotic cells in haematoxylin and eosin with that in crystal violet. 2To compare the number of mitotic figures present in normal oral mucosa, epithelial dysplasia and oral squamous cell carcinoma in crystal violet-stained sections with that in H and E-stained sections. Materials and Methods: Ten tissues of normal oral mucosa and 15 tissues each of oral epithelial dysplasia seen in tobacco-associated leukoplakia and squamous cell carcinoma were studied to evaluate the selectivity of 1% crystal violet for mitotic figures. The staining was compared with standard H and E staining. Statistical analysis was done using Man-Whitney U test. Results: A statistically significant increase in the mean mitotic count was observed in crystal violet-stained sections of epithelial dysplasia as compared to the H and E-stained sections ( p = 0.0327. A similar increase in the mitotic counts was noted in crystal violet-stained sections of oral squamous cell carcinoma as compared to the H and E-stained sections.( p = 0.0443. No significant difference was found in the mitotic counts determined in dysplasia or carcinoma by either the crystal violet ( p = 0.4429 or the H and E-staining techniques ( p = 0

Probing bacterial adhesion at the single-cell level

DEFF Research Database (Denmark)

Zeng, Guanghong; Müller, Torsten; Meyer, Rikke Louise

be considered. We have developed a simple and versatile method to make single-cell bacterial probes for measuring single cell adhesion by force spectroscopy using atomic force microscopy (AFM). A single-cell probe was readily made by picking up a bacterial cell from a glass surface by approaching a tipless AFM...... cantilever coated with the commercial cell adhesive CellTakTM. We applied the method to study adhesion of living cells to abiotic surfaces at the single-cell level. Immobilisation of single bacterial cells to the cantilever was stable for several hours, and viability was confirmed by Live/Dead staining...... on the adhesion force, we explored the bond formation and adhesive strength of four different bacterial strains towards three abiotic substrates with variable hydrophobicity and surface roughness. The adhesion force and final rupture length were dependent on bacterial strains, surfaces properties, and time...

A novel washing algorithm for underarm stain removal

Science.gov (United States)

Acikgoz Tufan, H.; Gocek, I.; Sahin, U. K.; Erdem, I.

2017-10-01

After contacting with human sweat which comprise around 27% sebum, anti-perspirants comprising aluminium chloride or its compounds form a jel-like structure whose solubility in water is very poor. In daily use, this jel-like structure closes sweat pores and hinders wetting of skin by sweat. However, when in contact with garments, they form yellowish stains at the underarm of the garments. These stains are very hard to remove with regular machine washing. In this study, first of all, we focused on understanding and simulating such stain formation on the garments. Two alternative procedures are offered to form jel-like structures. On both procedures, commercially available spray or deo-stick type anti-perspirants, standard acidic and basic sweat solutions and artificial sebum are used to form jel-like structures, and they are applied on fabric in order to get hard stains. Secondly, after simulation of the stain on the fabric, we put our efforts on developing a washing algorithm specifically designed for removal of underarm stains. Eight alternative washing algorithms are offered with varying washing temperature, amounts of detergent, and pre-stain removal procedures. Better algorithm is selected by comparison of Tristimulus Y values after washing.

Laser treatment of Port-wine stains

OpenAIRE

Boffa, Michael J.

2001-01-01

A state-of-the-art pulsed dye laser machine to treat port-wine stains and other vascular lesions has been available in the Malta Health Service since 1999. This article reviews the pathophysiology and clinical features of port- wine stains and describes the principles of laser treatment for this condition.

7 CFR 28.442 - Middling Yellow Stained Color.

Science.gov (United States)

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442... Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is deeper than Middling Tinged Color. [57 FR 34498, Aug. 5, 1992] below color grade cotton ...

Short Nissl staining for incubated cryostat sections of the brain.

Science.gov (United States)

Lindroos, O F

1991-01-01

Nissl stain often binds poorly to cryostat sections which have been incubated in solutions of radiolabeled ligands. Such incubation is used in receptor autoradiography of the brain when using the in vitro method. We have developed a rapid (16 min) modification of Nissl staining for sections that bind stain poorly, e.g., incubated sections. The method stains well sections which cannot be stained with other rapid Nissl staining methods.

Near-UV laser treatment of extrinsic dental enamel stains.

Science.gov (United States)

Schoenly, J E; Seka, W; Featherstone, J D B; Rechmann, P

2012-04-01

The selective ablation of extrinsic dental enamel stains using a 400-nm laser is evaluated at several fluences for completely removing stains with minimal damage to the underlying enamel. A frequency-doubled Ti:sapphire laser (400-nm wavelength, 60-nanosecond pulse duration, 10-Hz repetition rate) was used to treat 10 extracted human teeth with extrinsic enamel staining. Each tooth was irradiated perpendicular to the surface in a back-and-forth motion over a 1-mm length using an ∼300-µm-diam 10th-order super-Gaussian beam with fluences ranging from 0.8 to 6.4 J/cm(2) . Laser triangulation determined stain depth and volume removed by measuring 3D surface images before and after irradiation. Scanning electron microscopy evaluated the surface roughness of enamel following stain removal. Fluorescence spectroscopy measured spectra of unbleached and photobleached stains in the spectral range of 600-800 nm. Extrinsic enamel stains are removed with laser fluences between 0.8 and 6.4 J/cm(2) . Stains removed on sound enamel leave behind a smooth enamel surface. Stain removal in areas with signs of earlier cariogenic acid attacks resulted in isolated and randomly located laser-induced, 50-µm-diam enamel pits. These pits contain 0.5-µm diam, smooth craters indicative of heat transfer from the stain to the enamel and subsequent melting and water droplet ejection. Ablation stalling of enamel stains is typically observed at low fluences (Laser ablation of extrinsic enamel stains at 400 nm is observed to be most efficient above 3 J/cm(2) with minimal damage to the underlying enamel. Unsound underlying enamel is also observed to be selectively removed after irradiation. Copyright © 2012 Wiley Periodicals, Inc.

Port-wine stain

Science.gov (United States)

... About MedlinePlus Show Search Search MedlinePlus GO GO About MedlinePlus Site Map FAQs Customer Support Health Topics Drugs & Supplements Videos & Tools Español You Are Here: Home → Medical Encyclopedia → Port-wine stain URL of this page: //medlineplus.gov/ency/ ...

Effect of Melamine Sponge on Tooth Stain Removal.

Science.gov (United States)

Otsuka, Takero; Kawata, Toshitsugu

2015-01-01

To investigate the stain removal ability of melamine sponge before aesthetic tooth whitening in extracted teeth. Melamine sponge of thickness 40 mm was compressed and the destruction of the partition wall structure during the compression process was examined under a stereoscopic microscope. An extracted human tooth was cleaned by normal polishing or with melamine sponge for 90 s. To evaluate the stain level, the tooth surfaces were photographed under a stereoscopic microscope at 0, 30, 60 and 90 s. The residual stained region was traced in a high-magnification photograph, and the stain intensity was presented as a change, relative to the intensity before the experiment (0 s). Mechanical cleaning by toothbrushing produced polishing scratches on the tooth surface, whereas use of the melamine sponge resulted in only minimal scratches. As the compression level increased, the stain-removing effect tended to become stronger. Melamine sponge can remove stains from the tooth surface more effectively and less invasively compared to a conventional toothbrush. As no new scratches are made on the tooth surface when using a melamine sponge brush, the risk of re-staining is reduced. Cleaning using a melamine sponge brush can be easily and effectively performed at home and in a dental office.

LANTHANUM STAINING OF THE SURFACE COAT OF CELLS

Science.gov (United States)

Shea, Stephen M.

1971-01-01

Among the techniques which have been reported to stain the surface coat of cells, for electron microscopy, is lanthanum staining en bloc. Similarly, the presence of the cationic dye, Alcian blue 8GX, in a primary glutaraldehyde fixative has been reported to improve the preservation of the surface coat of cells of many types; however, the preserved coat is not very electron opaque unless thin sections are counterstained. The present paper shows that for several rat tissues lanthanum staining en bloc is an effective electron stain for the cell surface, giving excellent contrast, if combined sequentially with prefixation in an aldehyde fixative containing Alcian blue. The cationic substance cetylpyridinium chloride was found to have a similar effect to that of Alcian blue in enhancing the lanthanum staining of the surface coat material of the brush border of intestinal epithelial cells. The patterns of lanthanum staining obtained for the tissues studied strikingly resemble those reported in the literature where tissues are stained by several standard methods for demonstrating mucosubstances at the ultrastructural level. This fact and the reproduction of the effect of Alcian blue by cetylpyridinium chloride constitute a persuasive empirical argument that the material visualized is a mucopolysaccharide or mucopolysaccharide-protein complex. PMID:4108476

Fungal Fluorescence in Hematoxylin-Eosin Stained Sections

Directory of Open Access Journals (Sweden)

Murat Durdu

2017-06-01

Full Text Available A forty-six-year-old male presented to our dermatology clinic with two-year history of itching on his groin. His medical history revealed various topical corticosteroid creams without improvement of the skin lesion. Dermatological examination revealed erythematous nodules and follicular pustules on erythematous background on the inguinal area (Figure 1a. Potassium hydroxide (KOH examination was negative. Tzanck smear revealed abundant neutrophils without bacteria, fungi, or parasite. The histopathological examination showed granuloma formation with multinuclear giant cells and Periodic acid-Schiff (PAS-positive hyphae and spores around the hair follicles (Figure 1b, 1c. Hematoxylin-eosin (H&E-stained slides were examined under an immunofluorescence microscope, and these hyphae and spores showed autofluorescence (Figure 1d. Based on the clinical and histopathological findings, a Majocchi’s granuloma was considered. All lesions disappeared with topical and systemic terbinafine (250 mg/day treatment for six weeks.\tPearls;\tClinical: Not only bacteria, but also fungi, parasites, and viruses may cause folliculitis. Cytology should be initially done to identify the causes of infectious folliculitis. In case of negative cytology, histopathological examination and molecular methods can be used.\tCytological: To cytologically identify all of the causes of folliculitis, four separate samples should be taken: (i the first sample is stained with May-Grünwald-Giemsa for routine cytological examination; (ii the second sample is used for KOH testing; (iii the third sample is stained with an acid-fast stain to detect mycobacteria; and (iv the last specimen is Gram-stained to identify whether it is Gram-positive or Gram-negative (1. Histopathological: In infectious diseases, a definitive diagnosis should be done to identify the etiologic agent. The detection of fungal elements is challenging, when histopathological examination is performed with the H

Modified Alizarin Red S-Alcian Blue Staining for Reptilian Skeleton

Directory of Open Access Journals (Sweden)

Muhammad Ja’far Luthfi

2016-04-01

Full Text Available Skeletal staining is an important method in anatomical study. The aim of the research was to develop staining and clearing method of Reptilian skeleton using Alizarin Red S-Alcian Blue. The specimen were eviscerated, fixed, stained, cleared, and keep in glycerine solution. This modified double-staining has successfully stain bone and cartilage of Reptilian.

Imaging of oxygenation in 3D tissue models with multi-modal phosphorescent probes

Science.gov (United States)

Papkovsky, Dmitri B.; Dmitriev, Ruslan I.; Borisov, Sergei

2015-03-01

Cell-penetrating phosphorescence based probes allow real-time, high-resolution imaging of O2 concentration in respiring cells and 3D tissue models. We have developed a panel of such probes, small molecule and nanoparticle structures, which have different spectral characteristics, cell penetrating and tissue staining behavior. The probes are compatible with conventional live cell imaging platforms and can be used in different detection modalities, including ratiometric intensity and PLIM (Phosphorescence Lifetime IMaging) under one- or two-photon excitation. Analytical performance of these probes and utility of the O2 imaging method have been demonstrated with different types of samples: 2D cell cultures, multi-cellular spheroids from cancer cell lines and primary neurons, excised slices from mouse brain, colon and bladder tissue, and live animals. They are particularly useful for hypoxia research, ex-vivo studies of tissue physiology, cell metabolism, cancer, inflammation, and multiplexing with many conventional fluorophors and markers of cellular function.

[Histochemical stains for minerals by hematoxylin-lake method].

Science.gov (United States)

Miyagawa, Makoto

2013-04-01

The present study was undertaken to establish the experimental animal model by histological staining methods for minerals. After intraperitoneal injections of minerals, precipitates deposited on the surface of the liver. Liver tissues were fixed in paraformaldehyde, embedded in paraffin and cut into thin sections which were used as minerals containing standard section. Several reagents for histological stains and spectrophotometry for minerals were applied in both test-tube experiments and stainings of tissue sections to test for minerals. Hematoxylin-lake was found of capable of staining minerals in tissue. A simple technique used was described for light microscopic detection of minerals.

A fluorescence in situ staining method for investigating spores and vegetative cells of Clostridia by confocal laser scanning microscopy and structured illuminated microscopy.

Science.gov (United States)

D'Incecco, P; Ong, L; Gras, S; Pellegrino, L

2018-04-18

Non-pathogenic spore-forming Clostridia are of increasing interest due to their application in biogas production and their capability to spoil different food products. The life cycle for Clostridium includes a spore stage that can assist in survival under environmentally stressful conditions, such as extremes of temperature or pH. Due to their size, spores can be investigated by a range of microscopic techniques, many of which involve sample pre-treatment. We have developed a quick, simple and non-destructive fluorescent staining procedure that allows a clear differentiation between spores and vegetative cells and effectively stains spores, allowing recovery and tracking in subsequent experiments. Hoechst 34580, Propidium iodide and wheat germ agglutinin WGA 488 were used in combination to stain four strains of Clostridia at different life cycle stages. Staining was conducted without drying the sample, preventing changes induced by dehydration and cells observed by confocal laser scanner microscopy or using a super-resolution microscope equipped with a 3D-structured illumination module. Dual staining with Hoechst/Propidium iodide differentiated spores from vegetative cells, provided information on the viability of cells and was successfully applied to follow spore production induced by heating. Super-resolution microscopy of spores probed by Hoechst 34580 also allowed chromatin to be visualised. Direct staining of a cheese specimen using Nile Red and Fast Green allowed in situ observation of spores within the cheese and their position within the cheese matrix. The proposed staining method has broad applicability and can potentially be applied to follow Clostridium spore behaviour in a range of different environments. Copyright © 2018 Elsevier Ltd. All rights reserved.

Stool Gram stain

Science.gov (United States)

... stool sample. The Gram stain method is sometimes used to quickly diagnose bacterial infections. How the Test is Performed You will need to collect a stool sample. There are many ways to collect the sample. You can catch the stool on plastic wrap that is loosely placed over the toilet bowl ...

General Staining and Segmentation Procedures for High Content Imaging and Analysis.

Science.gov (United States)

Chambers, Kevin M; Mandavilli, Bhaskar S; Dolman, Nick J; Janes, Michael S

2018-01-01

Automated quantitative fluorescence microscopy, also known as high content imaging (HCI), is a rapidly growing analytical approach in cell biology. Because automated image analysis relies heavily on robust demarcation of cells and subcellular regions, reliable methods for labeling cells is a critical component of the HCI workflow. Labeling of cells for image segmentation is typically performed with fluorescent probes that bind DNA for nuclear-based cell demarcation or with those which react with proteins for image analysis based on whole cell staining. These reagents, along with instrument and software settings, play an important role in the successful segmentation of cells in a population for automated and quantitative image analysis. In this chapter, we describe standard procedures for labeling and image segmentation in both live and fixed cell samples. The chapter will also provide troubleshooting guidelines for some of the common problems associated with these aspects of HCI.

A novel acidic pH fluorescent probe based on a benzothiazole derivative

Science.gov (United States)

Ma, Qiujuan; Li, Xian; Feng, Suxiang; Liang, Beibei; Zhou, Tiqiang; Xu, Min; Ma, Zhuoyi

2017-04-01

A novel acidic pH fluorescent probe 1 based on a benzothiazole derivative has been designed, synthesized and developed. The linear response range covers the acidic pH range from 3.44 to 6.46, which is valuable for pH researches in acidic environment. The evaluated pKa value of the probe 1 is 4.23. The fluorescence enhancement of the studied probe 1 with an increase in hydrogen ions concentration is based on the hindering of enhanced photo-induced electron transfer (PET) process. Moreover, the pH sensor possesses a highly selective response to H+ in the presence of metal ions, anions and other bioactive small molecules which would be interfere with its fluorescent pH response. Furthermore, the probe 1 responds to acidic pH with short response time that was less than 1 min. The probe 1 has been successfully applied to confocal fluorescence imaging in live HeLa cells and can selectively stain lysosomes. All of such good properties prove it can be used to monitoring pH fluctuations in acidic environment with high sensitivity, pH dependence and short response time.

Comparism of Various Staining Techniques in the Diagnosis of ...

African Journals Online (AJOL)

SITWALA COMPUTERS

external intermediate host, usually an animal, in which sporogenesis and oocyst ... the parasite was detected in 111 of the samples stained,. 100(90.0%) of which .... screen stained slide was the auramine fluorochrome stain. The widely used ...

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

Science.gov (United States)

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

2009-01-01

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

[Usefulness of sputum Gram staining in community-acquired pneumonia].

Science.gov (United States)

Sato, Tadashi; Aoshima, Masahiro; Ohmagari, Norio; Tada, Hiroshi; Chohnabayashi, Naohiko

2002-07-01

To evaluate the usefulness of sputum gram staining in community-acquired pneumonia (CAP), we reviewed 144 cases requiring hospitalization in the last 4 years. The sensitivity was 75.5%, specificity 68.2%, positive predictive value 74.1%, negative predictive value 69.8%, positive likelihood ratio 2.37, negative likelihood ratio 0.36 and accuracy 72.2% in 97 cases. Both sputum gram staining and culture were performed. Concerning bacterial pneumonia (65 cases), we compared the Gram staining group (n = 33), which received initial antibiotic treatment, based on sputum gram staining with the Empiric group (n = 32) that received antibiotics empirically. The success rates of the initial antibiotic treatment were 87.9% vs. 78.1% (P = 0.473); mean hospitalization periods were 9.67 vs. 11.75 days (P = 0.053); and periods of intravenous therapy were 6.73 vs. 7.91 days (P = 0.044), respectively. As for initial treatment, penicillins were used in the Gram staining group more frequently (P gram staining is useful for the shortening of the treatment period and the appropriate selection of initial antibiotics in bacterial pneumonia. We believe, therefore, that sputum gram staining is indispensable as a diagnostic tool CAP.

Gram staining for the treatment of peritonsillar abscess.

Science.gov (United States)

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

2012-01-01

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

7 CFR 28.441 - Strict Middling Yellow Stained Color.

Science.gov (United States)

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Middling Yellow Stained Color. 28.441 Section... Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of Strict Middling Tinged Color. [57 FR 34498, Aug. 5, 1992] ...

CD3 immunohistochemical staining in diagnosis of lymphocytic colitis

DEFF Research Database (Denmark)

Fiehn, Anne-Marie Kanstrup; Engel, Ulla; Holck, Susanne

2016-01-01

and eosin (HE) stainings were available. At the second assessment, a supplementary CD3 immunohistochemical staining was also available. The aim was to evaluate whether a supplementary CD3 would increase the diagnostic agreement among pathologists, and whether a CD3 stain would change the diagnosis based...

Gram staining in the diagnosis of acute septic arthritis.

Science.gov (United States)

Faraj, A A; Omonbude, O D; Godwin, P

2002-10-01

This study aimed at determining the sensitivity and specificity of Gram staining of synovial fluid as a diagnostic tool in acute septic arthritis. A retrospective study was made of 22 patients who had arthroscopic lavage following a provisional diagnosis of acute septic arthritis of the knee joint. Gram stains and cultures of the knee aspirates were compared with the clinical and laboratory parameters, to evaluate their usefulness in diagnosing acute arthritis. All patients who had septic arthritis had pain, swelling and limitation of movement. CRP was elevated in 90% of patients. The incidence of elevated white blood cell count was higher in the group of patients with a positive Gram stain study (60%) as compared to patients with a negative Gram stain study (33%). Gram staining sensitivity was 45%. Its specificity was however 100%. Gram staining is an unreliable tool in early decision making in patients requiring urgent surgical drainage and washout.

Post-staining electroblotting for efficient and reliable peptide blotting.

Science.gov (United States)

Lee, Der-Yen; Chang, Geen-Dong

2015-01-01

Post-staining electroblotting has been previously described to transfer Coomassie blue-stained proteins from polyacrylamide gel onto polyvinylidene difluoride (PVDF) membranes. Actually, stained peptides can also be efficiently and reliably transferred. Because of selective staining procedures for peptides and increased retention of stained peptides on the membrane, even peptides with molecular masses less than 2 kDa such as bacitracin and granuliberin R are transferred with satisfactory results. For comparison, post-staining electroblotting is about 16-fold more sensitive than the conventional electroblotting for visualization of insulin on the membrane. Therefore, the peptide blots become practicable and more accessible to further applications, e.g., blot overlay detection or immunoblotting analysis. In addition, the efficiency of peptide transfer is favorable for N-terminal sequence analysis. With this method, peptide blotting can be normalized for further analysis such as blot overlay assay, immunoblotting, and N-terminal sequencing for identification of peptide in crude or partially purified samples.

Western Blot of Stained Proteins from Dried Polyacrylamide Gels

Science.gov (United States)

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.

Improved method for combination of immunocytochemistry and Nissl staining.

Science.gov (United States)

Kádár, Andrea; Wittmann, Gábor; Liposits, Zsolt; Fekete, Csaba

2009-10-30

Nissl staining is a widely used method to study morphology and pathology of neural tissue. After standard immunocytochemistry, the Nissl staining labels only the nucleus of neurons and the characteristic staining of the neuronal perikarya is absent or very weak. We hypothesized that the RNA degradation during the immunocytochemical treatment results in the loss of cytoplasmic staining with Nissl-dyes. To test this hypothesis, we used RNAse-free conditions for all steps of immunostaining. To further prevent the RNA-degradation by RNAse contaminations, the RNAse inhibitor heparin was added to all antibody-containing solutions. The efficiency of Nissl staining after standard and RNAse-free double-labeling immunocytochemistry was compared using antibodies against c-Fos and neuropeptide Y (NPY) on tissues of rats refed after 3 days of fasting. After standard immunocytochemistry, the Nissl-staining labeled the nuclei of neurons and only very faintly the cytoplasm of these cells. The RNAse-free treatment did not alter the distribution of immunoreaction signal, but preserved the staining of neuronal perikarya by the Nissl-dyes. In conclusion, the RNAse-free conditions during immunocytochemistry allow the labeling of neuronal perikarya by Nissl-dyes. The described method facilitates the mapping of immunocytochemical signals and makes possible the light microscopic examination of the innervation of neurons identified by their nuclear protein content.

Silver and Cyanine Staining of Oligonucleotides in Polyacrylamide Gel.

Science.gov (United States)

Tang, Weizhong; Zhou, Huafu; Li, Wei

2015-01-01

To explore why some oligonucleotides in denaturing polyacrylamide gel could not be silver-stained, 134 different oligonucleotides were analyzed using denaturing polyacrylamide gel electrophoresis stained with silver and asymmetric cyanine. As a result, we found that the sensitivity of oligos (dA), (dC), (dG) and (dT) to silver staining could be ranged as (dA) > (dG) > (dC) > (dT) from high to low. It was unexpected that oligo (dT) was hard to be silver-stained. Moreover, the silver staining of an oligonucleotide containing base T could be partially or completely inhibited by base T. The inhibition of silver staining by base T was a competitive inhibition which could be affected by the amounts of the argyrophil nucleobase and base T, the cis-distance between the argyrophil nucleobase and base T, and the gel concentration. The changes of the intensity of an oligonucleotide band caused by the changes of DNA base composition were diverse and interesting. The intensity of some oligonucleotide bands would significantly change when the changes of DNA base composition accumulated to a certain extent (usually ≥ 4 nt). The sensitivity of cyanine staining of ≤ 11-nt long oligonucleotides could be enhanced about 250-fold by fixing the gels with methanol fixing solution.

Maternal and fetal characteristics associated with meconium-stained amniotic fluid

DEFF Research Database (Denmark)

Balchin, Imelda; Whittaker, John C; Lamont, Ronald F

2011-01-01

To estimate the rates of meconium-stained amniotic fluid (AF) and adverse outcome in relation to gestational age and racial group, and to investigate the predictors of meconium-stained AF.......To estimate the rates of meconium-stained amniotic fluid (AF) and adverse outcome in relation to gestational age and racial group, and to investigate the predictors of meconium-stained AF....

Techniques for controlling variability in gram staining of obligate anaerobes.

Science.gov (United States)

Johnson, M J; Thatcher, E; Cox, M E

1995-01-01

Identification of anaerobes recovered from clinical samples is complicated by the fact that certain gram-positive anaerobes routinely stain gram negative; Peptostreptococcus asaccharolyticus, Eubacterium plautii, Clostridium ramosum, Clostridium symbiosum, and Clostridium clostridiiforme are among the nonconformists with regard to conventional Gram-staining procedures. Accurate Gram staining of American Type Culture Collection strains of these anaerobic bacteria is possible by implementing fixing and staining techniques within a gloveless anaerobic chamber. Under anaerobic conditions, gram-positive staining occurred in all test organisms with "quick" fixing techniques with both absolute methanol and formalin. The results support the hypothesis that, when anaerobic bacteria are exposed to oxygen, a breakdown of the physical integrity of the cell wall occurs, introducing Gram stain variability in gram-positive anaerobes. PMID:7538512

Methyl green-pyronin Y staining of nucleic acids: studies on the effects of staining time, dye composition and diffusion rates

DEFF Research Database (Denmark)

Prentø, P; Lyon, H O

2003-01-01

individually, simultaneously and sequentially. The results are presented as color charts approximating the observed staining patterns using a computerized palette. Our results indicate unequivocally that the differential staining is not time-dependent, but that it is dictated by the relative concentrations...

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

Science.gov (United States)

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.

Nucleic Acid Amplification Testing and Sequencing Combined with Acid-Fast Staining in Needle Biopsy Lung Tissues for the Diagnosis of Smear-Negative Pulmonary Tuberculosis.

Science.gov (United States)

Jiang, Faming; Huang, Weiwei; Wang, Ye; Tian, Panwen; Chen, Xuerong; Liang, Zongan

2016-01-01

Smear-negative pulmonary tuberculosis (PTB) is common and difficult to diagnose. In this study, we investigated the diagnostic value of nucleic acid amplification testing and sequencing combined with acid-fast bacteria (AFB) staining of needle biopsy lung tissues for patients with suspected smear-negative PTB. Patients with suspected smear-negative PTB who underwent percutaneous transthoracic needle biopsy between May 1, 2012, and June 30, 2015, were enrolled in this retrospective study. Patients with AFB in sputum smears were excluded. All lung biopsy specimens were fixed in formalin, embedded in paraffin, and subjected to acid-fast staining and tuberculous polymerase chain reaction (TB-PCR). For patients with positive AFB and negative TB-PCR results in lung tissues, probe assays and 16S rRNA sequencing were used for identification of nontuberculous mycobacteria (NTM). The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy of PCR and AFB staining were calculated separately and in combination. Among the 220 eligible patients, 133 were diagnosed with TB (men/women: 76/57; age range: 17-80 years, confirmed TB: 9, probable TB: 124). Forty-eight patients who were diagnosed with other specific diseases were assigned as negative controls, and 39 patients with indeterminate final diagnosis were excluded from statistical analysis. The sensitivity, specificity, PPV, NPV, and accuracy of histological AFB (HAFB) for the diagnosis of smear-negative were 61.7% (82/133), 100% (48/48), 100% (82/82), 48.5% (48/181), and 71.8% (130/181), respectively. The sensitivity, specificity, PPV, and NPV of histological PCR were 89.5% (119/133), 95.8% (46/48), 98.3% (119/121), and 76.7% (46/60), respectively, demonstrating that histological PCR had significantly higher accuracy (91.2% [165/181]) than histological acid-fast staining (71.8% [130/181]), P pulmonary tuberculosis. For patients with positive histological AFB and

Tumor Budding Detection by Immunohistochemical Staining is Not Superior to Hematoxylin and Eosin Staining for Predicting Lymph Node Metastasis in pT1 Colorectal Cancer.

Science.gov (United States)

Okamura, Takuma; Shimada, Yoshifumi; Nogami, Hitoshi; Kameyama, Hitoshi; Kobayashi, Takashi; Kosugi, Shin-ichi; Wakai, Toshifumi; Ajioka, Yoichi

2016-05-01

Tumor budding is recognized as an important risk factor for lymph node metastasis in pT1 colorectal cancer. Immunohistochemical staining for cytokeratin has the potential to improve the objective diagnosis of tumor budding over detection based on hematoxylin and eosin staining. However, it remains unclear whether tumor budding detected by immunohistochemical staining is a significant predictor of lymph node metastasis in pT1 colorectal cancer. The purpose of this study was to clarify the clinical significance of tumor budding detected by immunohistochemical staining in comparison with that detected by hematoxylin and eosin staining. This was a retrospective study. The study was conducted at Niigata University Medical & Dental Hospital. We enrolled 265 patients with pT1 colorectal cancer who underwent surgery with lymph node dissection. Tumor budding was evaluated by both hematoxylin and eosin and immunohistochemical staining with the use of CAM5.2 antibody. Receiver operating characteristic curve analyses were conducted to determine the optimal cutoff values for tumor budding detected by hematoxylin and eosin and CAM5.2 staining. Univariate and multivariate analyses were performed to identify the significant factors for predicting lymph node metastasis. Receiver operating characteristic curve analyses revealed that the cutoff values for tumor budding detected by hematoxylin and eosin and CAM5.2 staining for predicting lymph node metastases were 5 and 8. On multivariate analysis, histopathological differentiation (OR, 6.21; 95% CI, 1.16-33.33; p = 0.03) and tumor budding detected by hematoxylin and eosin staining (OR, 4.91; 95% CI, 1.64-14.66; p = 0.004) were significant predictors for lymph node metastasis; however, tumor budding detected by CAM5.2 staining was not a significant predictor. This study was limited by potential selection bias because surgically resected specimens were collected instead of endoscopically resected specimens. Tumor budding detected by

MANAJEMEN SARANA DAN PRASARANA PENDIDIKAN DI STAIN PAMEKASAN

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M. Muchlis Solichin

2011-07-01

Full Text Available Mediums management and pre-mediums represent an absolute done in an higher education institute, because Mediums and premediums in education management represent the absolut condition in the effort to reach the target which is expected. Thereby, Every the education organizer have to pay attention and conscripting the mind and energy to carry out education management that is professional and fulfill Standard National Education ( SNP. This Research copes to comprehend the mediums and pre-mediums management of education in STAIN Pamekasan, because during this time of mediums and basic mediums management are not yet showing its idealitas. This research is focussed at; a How mediums and pre-mediums menegement in STAIN Pamekasan ?,and b what Factors influencing mediums and pre-mediums management in STAIN Pamekasan ?. This research uses the qualitative type by using observation, interview, and documentation method. Based the rearch done, to be expressed that the first of STAIN Pamekasan conduct mediums and pre-mediums manegement still have the centralization character of top down, either in the case of planning, organizational, observation, and assessment of mediums and pre-mediums management owned, second in some cases of STAIN Pamekasan do not yet manage the mediums and pre-mediums management because they are caused by factor is its lack of management professionalism, either when doing the planning, organizational, treatment and observation or evaluation. Based the matter above, hence, suggested that STAIN Pamekasan carry out the mediums and pre-mediums management of education professionally.

Mapping stain distribution in pathology slides using whole slide imaging

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Fang-Cheng Yeh

2014-01-01

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

Efficacy test of a toothpaste in reducing extrinsic dental stain

Science.gov (United States)

Agustanti, A.; Ramadhani, S. A.; Adiatman, M.; Rahardjo, A.; Callea, M.; Yavuz, I.; Maharani, D. A.

2017-08-01

This clinical trial compared the external dental stain reduction achieved by tested toothpaste versus placebo in adult patients. In this double-blind, parallel, randomised clinical trial, 45 female volunteers with a mean age of 20 years old were included. All study subjects front teeth were topically applicated with Silver Diamine Fluoride (SDF) to create external dental stains. Subjects were randomized into test (n=22) and control (n=23) groups. Toothpastes were used for two days to analyse the effects of removing external stains on the labial surfaces of all anterior teeth. VITA Easyshade Advance 4.0 was used to measure dental extrinsic stains changes. The analysis showed statistically significant efficacy of the tested toothpaste in reducing external dental stain caused by SDF, comparing to the placebo toothpaste, after one and two days of usage. The tested toothpaste was effective in reducing dental stain.

Factors influencing extract of Hibiscus sabdariffa staining of rat testes.

Science.gov (United States)

Bassey, R B; Bakare, A A; Peter, A I; Oremosu, A A; Osinubi, A A

2012-08-01

Some plant extracts can be used in biology and medicine to reveal or identify cellular components and tissues. We investigated the effects of time and concentration on staining of histological sections of rat testes by an acidified extract of Hibiscus sabdariffa. An ethanolic extract of H. sabdariffa was diluted using 1% acetic acid in 70% ethanol to stain histological sections of testes at concentrations of 0.2, 0.1 and 0.05 g/ml for 5, 10, 15, 30, 45 and 60 min. The sections of testes were stained deep red. The staining efficiency of H. sabdariffa was greater at a high concentration and required less time to achieve optimal staining. H. sabdariffa is a strongly basic dye that can be used for various diagnostic purposes. Staining time and concentration must be considered to achieve optimal results.

Utility of Gram staining for diagnosis of Malassezia folliculitis.

Science.gov (United States)

Tu, Wei-Ting; Chin, Szu-Ying; Chou, Chia-Lun; Hsu, Che-Yuan; Chen, Yu-Tsung; Liu, Donald; Lee, Woan-Ruoh; Shih, Yi-Hsien

2018-02-01

Malassezia folliculitis (MalF) mimics acne vulgaris and bacterial folliculitis in clinical presentations. The role of Gram staining in rapid diagnosis of MalF has not been well studied. In our study, 32 patients were included to investigate the utility of Gram staining for MalF diagnosis. The final diagnoses of MalF were determined according to clinical presentation, pathological result and treatment response to antifungal agents. Our results show that the sensitivity and specificity of Gram staining are 84.6% and 100%, respectively. In conclusion, Gram staining is a rapid, non-invasive, sensitive and specific method for MalF diagnosis. © 2017 Japanese Dermatological Association.

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

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Memarzadeh Mehrdad

2009-01-01

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

Nucleic Acid Amplification Testing and Sequencing Combined with Acid-Fast Staining in Needle Biopsy Lung Tissues for the Diagnosis of Smear-Negative Pulmonary Tuberculosis.

Directory of Open Access Journals (Sweden)

Faming Jiang

Full Text Available Smear-negative pulmonary tuberculosis (PTB is common and difficult to diagnose. In this study, we investigated the diagnostic value of nucleic acid amplification testing and sequencing combined with acid-fast bacteria (AFB staining of needle biopsy lung tissues for patients with suspected smear-negative PTB.Patients with suspected smear-negative PTB who underwent percutaneous transthoracic needle biopsy between May 1, 2012, and June 30, 2015, were enrolled in this retrospective study. Patients with AFB in sputum smears were excluded. All lung biopsy specimens were fixed in formalin, embedded in paraffin, and subjected to acid-fast staining and tuberculous polymerase chain reaction (TB-PCR. For patients with positive AFB and negative TB-PCR results in lung tissues, probe assays and 16S rRNA sequencing were used for identification of nontuberculous mycobacteria (NTM. The sensitivity, specificity, positive predictive value (PPV, negative predictive value (NPV, and diagnostic accuracy of PCR and AFB staining were calculated separately and in combination.Among the 220 eligible patients, 133 were diagnosed with TB (men/women: 76/57; age range: 17-80 years, confirmed TB: 9, probable TB: 124. Forty-eight patients who were diagnosed with other specific diseases were assigned as negative controls, and 39 patients with indeterminate final diagnosis were excluded from statistical analysis. The sensitivity, specificity, PPV, NPV, and accuracy of histological AFB (HAFB for the diagnosis of smear-negative were 61.7% (82/133, 100% (48/48, 100% (82/82, 48.5% (48/181, and 71.8% (130/181, respectively. The sensitivity, specificity, PPV, and NPV of histological PCR were 89.5% (119/133, 95.8% (46/48, 98.3% (119/121, and 76.7% (46/60, respectively, demonstrating that histological PCR had significantly higher accuracy (91.2% [165/181] than histological acid-fast staining (71.8% [130/181], P < 0.001. Parallel testing of histological AFB staining and PCR showed the

Alcian blue-stained particles in a eutrophic lake

DEFF Research Database (Denmark)

Worm, J.; Søndergaard, Morten

1998-01-01

We used a neutral solution of Alcian Blue to stain transparent particles in eutrophic Lake Frederiksborg Slotss0, Denmark. Alcian Blue-stained particles (ABSP) appeared to be similar to the so-called transparent exopolymer particles (TEP) identified with an acidic solution of Alcian Blue. Our...

A Novel Contrast Stain for the Rapid Diagnosis of Pityriasis Versicolor: A Comparison of Chicago Sky Blue 6B Stain, Potassium Hydroxide Mount and Culture.

Science.gov (United States)

Lodha, Nikita; Poojary, Shital Amin

2015-01-01

The mycological study of pityriasis versicolor is usually done by potassium hydroxide (KOH) mount and culture. However, KOH mount lacks a color contrast and requires a trained eye to interpret, while culture is difficult to perform, time consuming and has low sensitivity. Chicago Sky Blue 6B (CSB) is a new contrast stain that highlights the fungal hyphae and spores, blue against a purplish background. This study was done to compare the utility of a novel contrast stain (CSB stain) with KOH mount and culture. Skin scrapings from the lesions of 100 clinically diagnosed cases of P. versicolor were subjected to (1) KOH mount and CSB stain for direct microscopic examination and (2) culture using Sabouraud's dextrose agar. The statistical analysis of CSB stain and culture was done using KOH mount as the reference method, as it is the most commonly performed and practical diagnostic test available for P. versicolor. An interrater reliability analysis using the Cohen's Kappa statistic was performed to determine consistency (agreement) among the different modalities. Direct microscopy with CSB stain, KOH mount and mycological culture showed positive results in 98 (98%), 92 (92%) and 56 (56%) patients, respectively. Using KOH mount as the reference method, CSB stain had a sensitivity of 100% which was significantly higher than culture (60.9%). Statistically significant fair agreement was found between CSB stain and KOH mount (94% with κ=0.38, P < 0.001). Negligible agreement was found between CSB stain and culture (66%, κ=0.199, P = 0.001) as well as between KOH mount and culture (64%, κ=0.051, P = 0.107). CSB staining of skin scrapings is the most sensitive method for the diagnosis of pityriasis versicolor. Due to the distinct contrast provided by CSB, it is easy to perform, rapid and qualitatively superior to KOH mount.

A useful single-solution polychrome stain for plant material...Brook Cyte-Chrome I.

Science.gov (United States)

Stanley L Krugman; Julia F. Littlefield

1968-01-01

Fresh and chemically fixed sectioned plant material can be quickly stained by applying a Brook Cyte Chrome I polychrome stain. Staining time averaged only about 10 minutes. And exact timing of staining and de-staining is not as critical as with most of the commonly used stains. The overall quality is comparable to that of the traditional stains.

Nuclear staining with alum hematoxylin.

Science.gov (United States)

Llewellyn, B D

2009-08-01

The hematoxylin and eosin stain is the most common method used in anatomic pathology, yet it is a method about which technologists ask numerous questions. Hematoxylin is a natural dye obtained from a tree originally found in Central America, and is easily converted into the dye hematein. This dye forms coordination compounds with mordant metals, such as aluminum, and the resulting lake attaches to cell nuclei. Regressive formulations contain a higher concentration of dye than progressive formulations and may also contain a lower concentration of mordant. The presence of an acid increases the life of the solution and in progressive solutions may also affect selectivity of staining. An appendix lists more than 60 hemalum formulations and the ratio of dye to mordant for each.

Stained Glass and Flu

Centers for Disease Control (CDC) Podcasts

2017-02-01

Dr. Robert Webster, an Emeritus member of the Department of Infectious Diseases at St. Jude Children's Research Hospital, discusses his cover art story on stained glass and influenza.  Created: 2/1/2017 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 2/1/2017.

Clinical utility of an automated instrument for gram staining single slides.

Science.gov (United States)

Baron, Ellen Jo; Mix, Samantha; Moradi, Wais

2010-06-01

Gram stains of 87 different clinical samples were prepared by the laboratory's conventional methods (automated or manual) and by a new single-slide-type automated staining instrument, GG&B AGS-1000. Gram stains from either heat- or methanol-fixed slides stained with the new instrument were easy to interpret, and results were essentially the same as those from the methanol-fixed slides prepared as a part of the routine workflow. This instrument is well suited to a rapid-response laboratory where Gram stain requests are commonly received on a stat basis.

The diverse and dynamic nature of Leishmania parasitophorous vacuoles studied by multidimensional imaging.

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Fernando Real

Full Text Available An important area in the cell biology of intracellular parasitism is the customization of parasitophorous vacuoles (PVs by prokaryotic or eukaryotic intracellular microorganisms. We were curious to compare PV biogenesis in primary mouse bone marrow-derived macrophages exposed to carefully prepared amastigotes of either Leishmania major or L. amazonensis. While tight-fitting PVs are housing one or two L. major amastigotes, giant PVs are housing many L. amazonensis amastigotes. In this study, using multidimensional imaging of live cells, we compare and characterize the PV biogenesis/remodeling of macrophages i hosting amastigotes of either L. major or L. amazonensis and ii loaded with Lysotracker, a lysosomotropic fluorescent probe. Three dynamic features of Leishmania amastigote-hosting PVs are documented: they range from i entry of Lysotracker transients within tight-fitting, fission-prone L. major amastigote-housing PVs; ii the decrease in the number of macrophage acidic vesicles during the L. major PV fission or L. amazonensis PV enlargement; to iii the L. amazonensis PV remodeling after homotypic fusion. The high content information of multidimensional images allowed the updating of our understanding of the Leishmania species-specific differences in PV biogenesis/remodeling and could be useful for the study of other intracellular microorganisms.

News from the Biological Stain Commission No. 11

DEFF Research Database (Denmark)

Lyon, H O; Horobin, R W

2012-01-01

The 11th issue of News from the Biological Stain Commission (BSC) provides our first impressions of the REACH and ECHA programs. We intend to give a more thorough account of what these important programs actually mean in later editions of News from the Biological Stain Commission. Under the heading...

A low molecular weight zinc2+-dipicolylamine-based probe detects apoptosis during tumour treatment better than an annexin V-based probe

International Nuclear Information System (INIS)

Palmowski, Karin; Rix, Anne; Lederle, Wiltrud; Kiessling, Fabian; Behrendt, Florian F.; Mottaghy, Felix M.; Gray, Brian D.; Pak, Koon Y.; Palmowski, Moritz

2014-01-01

Molecular imaging of apoptosis is frequently discussed for monitoring cancer therapies. Here, we compare the low molecular weight phosphatidylserine-targeting ligand zinc 2+ -dipicolylamine (Zn 2+ -DPA) with the established but reasonably larger protein annexin V. Molecular apoptosis imaging with the fluorescently labelled probes annexin V (750 nm, 36 kDa) and Zn 2+ -DPA (794 nm, 1.84 kDa) was performed in tumour-bearing mice (A431). Three animal groups were investigated: untreated controls and treated tumours after 1 or 4 days of anti-angiogenic therapy (SU11248). Additionally, μPET with 18 F-FDG was performed. Imaging data were displayed as tumour-to-muscle ratio (TMR) and validated by quantitative immunohistochemistry. Compared with untreated control tumours, TUNEL staining indicated significant apoptosis after 1 day (P 2+ -DPA uptake increased significantly after 1 day (P 2+ -DPA, 18 F-FDG tumour uptake decreased significantly at days 1 (P 2+ -DPA than with the annexin V-based probe. Additionally, significant treatment effects were detectable as early using Zn 2+ -DPA as with measurements of the glucose metabolism using 18 F-FDG. (orig.)

A novel contrast stain for the rapid diagnosis of pityriasis versicolor: A comparison of Chicago Sky Blue 6B stain, potassium hydroxide mount and culture

Directory of Open Access Journals (Sweden)

Nikita Lodha

2015-01-01

Full Text Available Background: The mycological study of pityriasis versicolor is usually done by potassium hydroxide (KOH mount and culture. However, KOH mount lacks a color contrast and requires a trained eye to interpret, while culture is difficult to perform, time consuming and has low sensitivity. Chicago Sky Blue 6B (CSB is a new contrast stain that highlights the fungal hyphae and spores, blue against a purplish background. Aims and Objectives: This study was done to compare the utility of a novel contrast stain (CSB stain with KOH mount and culture. Materials and Methods: Skin scrapings from the lesions of 100 clinically diagnosed cases of P. versicolor were subjected to (1 KOH mount and CSB stain for direct microscopic examination and (2 culture using Sabouraud′s dextrose agar. The statistical analysis of CSB stain and culture was done using KOH mount as the reference method, as it is the most commonly performed and practical diagnostic test available for P. versicolor. An interrater reliability analysis using the Cohen′s Kappa statistic was performed to determine consistency (agreement among the different modalities. Observations and Results: Direct microscopy with CSB stain, KOH mount and mycological culture showed positive results in 98 (98%, 92 (92% and 56 (56% patients, respectively. Using KOH mount as the reference method, CSB stain had a sensitivity of 100% which was significantly higher than culture (60.9%. Statistically significant fair agreement was found between CSB stain and KOH mount (94% with κ=0.38, P < 0.001. Negligible agreement was found between CSB stain and culture (66%, κ=0.199, P = 0.001 as well as between KOH mount and culture (64%, κ=0.051, P = 0.107. Conclusion: CSB staining of skin scrapings is the most sensitive method for the diagnosis of pityriasis versicolor. Due to the distinct contrast provided by CSB, it is easy to perform, rapid and qualitatively superior to KOH mount.

Acetylcholinesterase and Nissl staining in the same histological section.

Science.gov (United States)

Shipley, M T; Ennis, M; Behbehani, M M

1989-12-18

Acetylcholinesterase (AChE) enzyme histochemistry and Nissl staining are commonly utilized in neural architectonic studies. However, the opaque reaction deposit produced by the most commonly used AChE histochemical methods is not compatible with satisfactory Nissl staining. As a result, precise correlation of AChE and Nissl staining necessitates time-consuming comparisons of adjacent sections which may have differential shrinkage. Here, we have modified the Koelle-Friedenwald histochemical reaction for AChE by omitting the final intensification steps. The modified reaction yields a non-opaque reaction product that is selectively visualized by darkfield illumination. This non-intensified darkfield AChE (NIDA) reaction allows clear visualization of Nissl staining in the same histological section. This combined AChE-Nissl method greatly facilitates detailed correlation of enzyme and cytoarchitectonic organization.

Should gram stains have a role in diagnosing hip arthroplasty infections?

Science.gov (United States)

Johnson, Aaron J; Zywiel, Michael G; Stroh, D Alex; Marker, David R; Mont, Michael A

2010-09-01

The utility of Gram stains in diagnosing periprosthetic infections following total hip arthroplasty has recently been questioned. Several studies report low sensitivity of the test, and its poor ability to either confirm or rule out infection in patients undergoing revision total hip arthroplasty. Despite this, many institutions including that of the senior author continue to perform Gram stains during revision total hip arthroplasty. We assessed the sensitivity, specificity, accuracy, and positive and negative predictive values of Gram stains from surgical-site samplings taken from procedures on patients with both infected and aseptic revision total hip arthroplasties. A review was performed on patients who underwent revision total hip arthroplasty between 2000 and 2007. Eighty-two Gram stains were performed on patients who had infected total hip arthroplasties and underwent revision procedures. Additionally, of the 410 revision total hip arthroplasties performed on patients who were confirmed infection-free, 120 Gram stains were performed. Patients were diagnosed as infected using multiple criteria at the time of surgery. Sensitivity, specificity, positive and negative predictive values, and accuracy were calculated from these Gram stain results. The Gram stain demonstrated a sensitivity and specificity of 9.8% and 100%, respectively. In this series, the Gram stain had a negative predictive value of 62%, a positive predictive value of 100%, and an accuracy of 63%. Gram stains obtained from surgical-site samples had poor sensitivity and poor negative predictive value. Based on these findings, as well as those of other authors, we believe that Gram stains should no longer be considered for diagnosing infections in revision total hip arthroplasty. Level III, diagnostic study. See Guidelines for Authors for a complete description of levels of evidence.

Standardization in biological staining. The influence of dye manufacturing

DEFF Research Database (Denmark)

Lyon, H

2000-01-01

not have been subjected to quality assessment either internally by the producer or vendor or externally by independent investigators or organizations such as the Biological Stain Commission. Concerted attempts at standardization in Europe are discussed. The latest results of this work, the European...... standard EN 12376, is presented. This standard is concerned with information supplied by the manufacturer with in vitro diagnostic reagents for biological staining. The standard has been prepared by a Working Group on Staining in Biology under Technical Committee 140, In Vitro Medical Devices...

Helicobacter pylori detection in chronic gastritis: a comparison of staining methods

International Nuclear Information System (INIS)

Ahmad, F.; Khan, I.

2011-01-01

Background: Helicobacter pylori is an important cause of chronic gastritis, gastric ulceration and gastric malignancies as gastric carcinoma and MALT lymphoma. Its definitive diagnosis is based on histopathology. Routine H and E stain is not very effective in its detection, immune-stains and fluorescent stains are costly. Need for simple cheap and sensitive stain has always been a topic of hot debate and extensive research. Method: paraffin embedded blocks of all adult patients diagnosed as chronic gastritis/gastric ulceration with no accompanying gastric pathology as hypertrophic gastropathys, and neoplasias were taken into study. Three sections of 4 micron were cut and stained with routine H and E, Giemsa, and Cresyl fast violet. Results: Total number of patients was 50. Out of these 37 (74%) were males and 13 (26%) were females. Mean age of the patients was 50.4 years. Thirty-four percent (34%) were positive in normal H and E stain, 68% were positive in Giemsa and 76% were positive in Cresyl fast violet. Conclusion: Cresyl fast violet is a good stain for diagnosis of H. pylori gastritis. (author)

Decreased mortality associated with prompt Gram staining of blood cultures.

Science.gov (United States)

Barenfanger, Joan; Graham, Donald R; Kolluri, Lavanya; Sangwan, Gaurav; Lawhorn, Jerry; Drake, Cheryl A; Verhulst, Steven J; Peterson, Ryan; Moja, Lauren B; Ertmoed, Matthew M; Moja, Ashley B; Shevlin, Douglas W; Vautrain, Robert; Callahan, Charles D

2008-12-01

Gram stains of positive blood cultures are the most important factor influencing appropriate therapy. The sooner appropriate therapy is initiated, the better. Therefore, it is reasonable to expect that the sooner Gram stains are performed, the better. To determine the value of timely Gram stains and whether improvement in Gram stain turnaround time (TAT) is feasible, we compared data for matched pairs of patients with cultures processed promptly ( or =1 hour TAT) and then monitored TAT by control charting.In 99 matched pairs, average difference in time to detection of positive blood cultures within a pair of patients was less than 0.1 hour. For the less than 1 hour TAT group, the average TAT and crude mortality were 0.1 hour and 10.1%, respectively; for the 1 hour or longer TAT group, they were 3.3 hours and 19.2%, respectively (P Gram stains.

A new technique for Gram staining paraffin-embedded tissue.

Science.gov (United States)

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

1979-01-01

Five techniques for Gram staining bacteria in paraffin sections were compared on serial sections of pulmonary tissues from eight bacteriological necropsies. Brown and Hopp's method was the most satisfactory for distinguishing Gram-positive and Gram-negative bacteria. However, this method cannot be recommended as the preparations were frequently overstained, and the Gram-negative bacteria were stained indistinctly. A modification of Brown and Hopps' method was developed which stains larger numbers of Gram-negative bacteria and differentiates well between different cell types and connective tissue, and there is no risk of overstaining. PMID:86548

Application of GelGreen™ in Cesium Chloride Density Gradients for DNA-Stable Isotope Probing Experiments.

Directory of Open Access Journals (Sweden)

Jingfeng Gao

Full Text Available In this study, GelGreen™ was investigated as a replacement for SYBR® Safe to stain DNA in cesium chloride (CsCl density gradients for DNA-stable isotope probing (SIP experiments. Using environmental DNA, the usage of GelGreen™ was optimized for sensitivity compared to SYBR® Safe, its optimal concentration, detection limit for environmental DNA and its application in environmental DNA-SIP assay. Results showed that GelGreen™ was more sensitive than SYBR® Safe, while the optimal dosage (15X concentration needed was approximately one-third of SYBR® Safe, suggesting that its sensitivity was three times more superior than SYBR® Safe. At these optimal parameters, the detection limit of GelGreen™-stained environmental DNA was as low as 0.2 μg, but the usage of 0.5 μg environmental DNA was recommended to produce a more consistent DNA band. In addition, a modified needle extraction procedure was developed to withdraw DNA effectively by fractionating CsCl density gradients into four or five fractions. The successful application of GelGreen™ staining with 13C-labeled DNA from enriched activated sludge suggests that this stain was an excellent alternative of SYBR® Safe in CsCl density gradients for DNA-SIP assays.

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

International Nuclear Information System (INIS)

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

2000-01-01

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

A flow-cytometric gram-staining technique for milk-associated bacteria.

Science.gov (United States)

Holm, Claus; Jespersen, Lene

2003-05-01

A Gram-staining technique combining staining with two fluorescent stains, Oregon Green-conjugated wheat germ agglutinin (WGA) and hexidium iodide (HI) followed by flow-cytometric detection is described. WGA stains gram-positive bacteria while HI binds to the DNA of all bacteria after permeabilization by EDTA and incubation at 50 degrees C for 15 min. For WGA to bind to gram-positive bacteria, a 3 M potassium chloride solution was found to give the highest fluorescence intensity. A total of 12 strains representing some of the predominant bacterial species in bulk tank milk and mixtures of these were stained and analyzed by flow cytometry. Overall, the staining method showed a clear differentiation between gram-positive and gram-negative bacterial populations. For stationary-stage cultures of seven gram-positive bacteria and five gram-negative bacteria, an average of 99% of the cells were correctly interpreted. The method was only slightly influenced by the growth phase of the bacteria or conditions such as freezing at -18 degrees C for 24 h. For any of these conditions, an average of at least 95% of the cells were correctly interpreted. When stationary-stage cultures were stored at 5 degrees C for 14 days, an average of 86% of the cells were correctly interpreted. The Gram-staining technique was applied to the flow cytometry analysis of bulk tank milk inoculated with Staphylococcus aureus and Escherichia coli. These results demonstrate that the technique is suitable for analyzing milk samples without precultivation.

Evaluation of Papanicolaou stain for studying micronuclei in buccal cells under field conditions.

Science.gov (United States)

Ayyad, Sohair B A; Israel, Ebenezer; El-Setouhy, Maged; Nasr, Ghada Radwan; Mohamed, Mostafa K; Loffredo, Christopher A

2006-01-01

To compare Papanicolaou (Pap) and May-Grünwald Giemsa (MGG) stain as 2 techniques for staining for buccal mucosal cells to detect micronuclei (MN) infield studies. Eighty cytologic smears (2 per individual) were taken from the buccal mucosa of 40 cigarette smokers recruited at a rural village in Egypt. Forty smears were stained with Pap stain and 40 with MGG stain. All were assessed for cellularity and scored for MN. Pap stain was faster and easier to process and transport in the field study than was MGG stain. Regarding MGG smears, bacteria and cell debris masked the MN as compared to Pap smears, in which the fixative destroyed the bacteria and made the cell boundaries clearly demarcated. Using Pap stain, MN were seen easily in transparent cytoplasm. Pap stain is the preferred method infield studies for scoring and detecting MN in cells of buccal mucosa.

Kinetics of bacterial fluorescence staining with 3,3'-diethylthiacyanine.

Science.gov (United States)

Thomas, Marlon S; Nuñez, Vicente; Upadhyayula, Srigokul; Zielins, Elizabeth R; Bao, Duoduo; Vasquez, Jacob M; Bahmani, Baharak; Vullev, Valentine I

2010-06-15

For more than a century, colorimetric and fluorescence staining have been the foundation of a broad range of key bioanalytical techniques. The dynamics of such staining processes, however, still remains largely unexplored. We investigated the kinetics of fluorescence staining of two gram-negative and two gram-positive species with 3,3'-diethylthiacyanine (THIA) iodide. An increase in the THIA fluorescence quantum yield, induced by the bacterial dye uptake, was the principal reason for the observed emission enhancement. The fluorescence quantum yield of THIA depended on the media viscosity and not on the media polarity, which suggested that the microenvironment of the dye molecules taken up by the cells was restrictive. The kinetics of fluorescence staining did not manifest a statistically significant dependence neither on the dye concentration, nor on the cell count. In the presence of surfactant additives, however, the fluorescence-enhancement kinetic patterns manifested species specificity with statistically significant discernibility.

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

International Nuclear Information System (INIS)

Van Keuren, M.L.; Goldman, D.; Merril, C.R.

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 irreversible for 3 H detection and moderate for 14 C detection. A silver stain based on photochemical methods had minimal quenching of 14 C detection and less of a quenching effect than the histological stain for 3 H detection. The 3 H quenching effect was partially reversible for the photochemical stain

Deep-Red Fluorescent Gold Nanoclusters for Nucleoli Staining: Real-Time Monitoring of the Nucleolar Dynamics in Reverse Transformation of Malignant Cells.

Science.gov (United States)

Wang, Xiaojuan; Wang, Yanan; He, Hua; Ma, Xiqi; Chen, Qi; Zhang, Shuai; Ge, Baosheng; Wang, Shengjie; Nau, Werner M; Huang, Fang

2017-05-31

Nucleoli are important subnuclear structures inside cells. We report novel fluorescent gold nanoclusters (K-AuNCs) that are able to stain the nucleoli selectively and make it possible to explore the nucleolar morphology with fluorescence imaging technique. This novel probe is prepared through an easy synthesis method by employing a tripeptide (Lys-Cys-Lys) as the surface ligand. The properties, including deep-red fluorescence emission (680 nm), large Stocks shift, broad excitation band, low cytotoxicity, and good photostability, endow this probe with potential for bioanalytical applications. Because of their small size and their positively charged surface, K-AuNCs are able to accumulate efficiently at the nucleolar regions and provide precise morphological information. K-AuNCs are also used to monitor the nucleolar dynamics along the reverse-transformation process of malignant cells, induced by the agonist of protein A, 8-chloro-cyclic adenosine monophosphate. This gives a novel approach for investigating the working mechanism of antitumor drugs.

Etika Berbusana Mahasiswa Stain Samarinda

Directory of Open Access Journals (Sweden)

Ida Suryani Wijaya

2012-06-01

Full Text Available Ethics is about behavior of human being, such as which one is right or wrong. The ethics is always affecting the human life. The ethics gives people orientation how he/she do manything every time every day. Islamic ethics consists of the way how someone interact each other; how someone should do or not to do, how to sit, how to walk, how to eat or drink, how to sleep, or how to get dressed. Al-Qur’an uses three terms to define about dressing, they are: libas, tsiyah, and sarahi. Dressing has a function as covering the body, as assessoris, as the way to do Islamic taqwa, and as an identiy. Dressing ethics of the female students of STAIN Samarinda has been regulated by the rector regulation No 19 of the year 2002 about relation and dressing ethics for the students of STAIN Samarinda.

A Comparison of Heat versus Methanol Fixation for Gram Staining Bacteria

Science.gov (United States)

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…

Sperm viability staining in ecology and evolution: potential pitfalls

DEFF Research Database (Denmark)

Holman, Luke

2009-01-01

The causes and consequences of variation in sperm quality, survival and ageing are active areas of research in ecology and evolution. In order to address these topics, many recent studies have measured sperm viability using fluorescent staining. Although sperm viability staining has produced a nu...

Preparation of colloidal gold for staining proteins electrotransferred onto nitrocellulose membranes.

Science.gov (United States)

Yamaguchi, K; Asakawa, H

1988-07-01

This paper describes a simple method of preparing colloidal gold for staining protein blots. Colloidal gold was prepared from 0.005 or 0.01% HAuCl4 by the addition of formalin as a reductant and potassium hydroxide. Staining of small cell carcinoma tissue extract blotted onto nitrocellulose membranes with this colloidal gold solution resulted in the appearance of a large number of clear wine-red bands. The sensitivity of gold staining was 60 times higher than that of Coomassie brilliant blue staining and almost comparable to that of silver staining of proteins in polyacrylamide gel. The sensitivity of this method was also satisfactory in comparison with that of enzyme immunoblotting. The colloidal gold prepared by this method is usable for routine work.

Color and dichroism of silver-stained glasses

International Nuclear Information System (INIS)

Molina, Gloria; Murcia, Sonia; Molera, Judit; Roldan, Clodoaldo; Crespo, Daniel; Pradell, Trinitat

2013-01-01

Yellow decorations in glasses have been produced since the beginning of the fourteenth century by incorporating metallic silver nanoparticles into the glass (from a few to some tens of nanometers). The optical response of the glass-particles composite is determined by the surface plasmon resonance absorption and scattering of the nanometric metallic particles. Generally, the same color is perceived in reflection and in transmission although dichroic effects are occasionally observed. As silver-stained glasses were designed to be observed in transmission, tuning the transmission color from yellow to red was of technological interest. The relationship between the color observed both in transmission and reflection and the composition and nanostructure of regular (yellow) and dichroic (yellow and red) silver stains from the Renaissance (late fifteenth and sixteenth century, respectively) is related to the presence of a layer (of about 10–20 μm thick) of metallic silver nanoparticles (from few to 100 nm in size). The correlation between the colors observed and the silver stain nanostructure is studied with particular emphasis on the origin of the dichroic behavior. The optical response is computed and compared to the experimental data. Differences in the synthesis parameters responsible for the colors and for the dichroic behavior of the silver stain glasses are proposed. This is essential for the replication of the glass pieces which are required as replacements in the restoration/conservation of the windows but is also of broader interest

Color and dichroism of silver-stained glasses

Energy Technology Data Exchange (ETDEWEB)

Molina, Gloria [Universitat Politecnica de Catalunya, Center for Research in NanoEngineering (Spain); Murcia, Sonia [Universidad de Valencia, Instituto de Ciencia de los Materiales (Spain); Molera, Judit [Universitat de Vic, GRTD, Escola Politecnica Superior (Spain); Roldan, Clodoaldo [Universidad de Valencia, Instituto de Ciencia de los Materiales (Spain); Crespo, Daniel; Pradell, Trinitat, E-mail: Trinitat.Pradell@upc.edu [Universitat Politecnica de Catalunya, Center for Research in NanoEngineering (Spain)

2013-09-15

Yellow decorations in glasses have been produced since the beginning of the fourteenth century by incorporating metallic silver nanoparticles into the glass (from a few to some tens of nanometers). The optical response of the glass-particles composite is determined by the surface plasmon resonance absorption and scattering of the nanometric metallic particles. Generally, the same color is perceived in reflection and in transmission although dichroic effects are occasionally observed. As silver-stained glasses were designed to be observed in transmission, tuning the transmission color from yellow to red was of technological interest. The relationship between the color observed both in transmission and reflection and the composition and nanostructure of regular (yellow) and dichroic (yellow and red) silver stains from the Renaissance (late fifteenth and sixteenth century, respectively) is related to the presence of a layer (of about 10-20 {mu}m thick) of metallic silver nanoparticles (from few to 100 nm in size). The correlation between the colors observed and the silver stain nanostructure is studied with particular emphasis on the origin of the dichroic behavior. The optical response is computed and compared to the experimental data. Differences in the synthesis parameters responsible for the colors and for the dichroic behavior of the silver stain glasses are proposed. This is essential for the replication of the glass pieces which are required as replacements in the restoration/conservation of the windows but is also of broader interest.

Clinical Utility of an Automated Instrument for Gram Staining Single Slides ▿

Science.gov (United States)

Baron, Ellen Jo; Mix, Samantha; Moradi, Wais

2010-01-01

Gram stains of 87 different clinical samples were prepared by the laboratory's conventional methods (automated or manual) and by a new single-slide-type automated staining instrument, GG&B AGS-1000. Gram stains from either heat- or methanol-fixed slides stained with the new instrument were easy to interpret, and results were essentially the same as those from the methanol-fixed slides prepared as a part of the routine workflow. This instrument is well suited to a rapid-response laboratory where Gram stain requests are commonly received on a stat basis. PMID:20410348

Touch cytology in diagnosing Helicobacter pylori: comparison of four staining methods.

Science.gov (United States)

Hashemi, M R; Rahnavardi, M; Bikdeli, B; Dehghani Zahedani, M; Iranmanesh, F

2008-06-01

Helicobacter pylori (Hp), a major cause of peptic ulcer disease and an important risk factor for gastric malignancy, can be diagnosed by several methods. Touch cytology (TC) of the gastric mucosa has been noted to give good results and has been found to be very simple, inexpensive and rapid. However, evidence regarding the accuracy of different staining methods of TC is lacking. The present study aims at defining the diagnostic accuracy of four different staining methods of TC. Biopsy specimens were taken from the antral mucosa of one hundred consecutive patients referred for upper gastrointestinal endoscopy (UGIE) for various indications. TC slides were processed by four staining methods: Wright, Giemsa, Papanicolaou and Gram. Rapid urease test (RUT) and histological examination of specimens were also performed. The same experienced pathologist evaluated the coded samples. A patient's Hp status was established by minimum concordance of the three tests, including histology, RUT, and 'Touch mean'. The latter was defined positive when at least three of the four TC staining methods were positive. Forty-six patients (46%) were positive for Hp according to Hp status. TC stained by Wright had excellent agreement with both histology (kappa = 0.80, P stained TC (88.89%) was significantly more specific than both Giemsa- (74.07%; P stained (70.37%; P stained TC can safely substitute for histology. However, when assessment for severity of mucosal damage or cell atypias is meant, histology cannot be neglected.

Enhanced fluorescence imaging of live cells by effective cytosolic delivery of probes.

Directory of Open Access Journals (Sweden)

Marzia Massignani

Full Text Available BACKGROUND: Microscopic techniques enable real-space imaging of complex biological events and processes. They have become an essential tool to confirm and complement hypotheses made by biomedical scientists and also allow the re-examination of existing models, hence influencing future investigations. Particularly imaging live cells is crucial for an improved understanding of dynamic biological processes, however hitherto live cell imaging has been limited by the necessity to introduce probes within a cell without altering its physiological and structural integrity. We demonstrate herein that this hurdle can be overcome by effective cytosolic delivery. PRINCIPAL FINDINGS: We show the delivery within several types of mammalian cells using nanometre-sized biomimetic polymer vesicles (a.k.a. polymersomes that offer both highly efficient cellular uptake and endolysomal escape capability without any effect on the cellular metabolic activity. Such biocompatible polymersomes can encapsulate various types of probes including cell membrane probes and nucleic acid probes as well as labelled nucleic acids, antibodies and quantum dots. SIGNIFICANCE: We show the delivery of sufficient quantities of probes to the cytosol, allowing sustained functional imaging of live cells over time periods of days to weeks. Finally the combination of such effective staining with three-dimensional imaging by confocal laser scanning microscopy allows cell imaging in complex three-dimensional environments under both mono-culture and co-culture conditions. Thus cell migration and proliferation can be studied in models that are much closer to the in vivo situation.

Use of a Novel Rover-mounted Fluorescence Imager and Fluorescent Probes to Detect Biological Material in the Atacama Desert in Daylight

Science.gov (United States)

Weinstein, S.; Pane, D.; Warren-Rhodes, K.; Cockell, C.; Ernst, L. A.; Minkley, E.; Fisher, G.; Emani, S.; Wettergreen, D. S.; Wagner, M.

2005-01-01

We have developed an imaging system, the Fluorescence Imager (FI), for detecting fluorescence signals from sparse microorganisms and biofilms during autonomous rover exploration. The fluorescence signals arise both from naturally occurring chromophores, such as chlorophyll of cyanobacteria and lichens, and from fluorescent probes applied to soil and rocks. Daylight imaging has been accomplished by a novel use of a high-powered flashlamp synchronized to a CCD camera. The fluorescent probes are cell permanent stains that have extremely low intrinsic fluorescence (quantum yields less than 0.01) and a large fluorescence enhancement (quantum yields greater than 0.4) when bound to the target. Each probe specifically targets either carbohydrates, proteins, nucleic acids or membrane lipids, the four classes of macromolecules found in terrestrial life. The intent of the probes is to interrogate the environment for surface and endolithic life forms.

Comparison of algorithms for blood stain detection applied to forensic hyperspectral imagery

Science.gov (United States)

Yang, Jie; Messinger, David W.; Mathew, Jobin J.; Dube, Roger R.

2016-05-01

Blood stains are among the most important types of evidence for forensic investigation. They contain valuable DNA information, and the pattern of the stains can suggest specifics about the nature of the violence that transpired at the scene. Early detection of blood stains is particularly important since the blood reacts physically and chemically with air and materials over time. Accurate identification of blood remnants, including regions that might have been intentionally cleaned, is an important aspect of forensic investigation. Hyperspectral imaging might be a potential method to detect blood stains because it is non-contact and provides substantial spectral information that can be used to identify regions in a scene with trace amounts of blood. The potential complexity of scenes in which such vast violence occurs can be high when the range of scene material types and conditions containing blood stains at a crime scene are considered. Some stains are hard to detect by the unaided eye, especially if a conscious effort to clean the scene has occurred (we refer to these as "latent" blood stains). In this paper we present the initial results of a study of the use of hyperspectral imaging algorithms for blood detection in complex scenes. We describe a hyperspectral imaging system which generates images covering 400 nm - 700 nm visible range with a spectral resolution of 10 nm. Three image sets of 31 wavelength bands were generated using this camera for a simulated indoor crime scene in which blood stains were placed on a T-shirt and walls. To detect blood stains in the scene, Principal Component Analysis (PCA), Subspace Reed Xiaoli Detection (SRXD), and Topological Anomaly Detection (TAD) algorithms were used. Comparison of the three hyperspectral image analysis techniques shows that TAD is most suitable for detecting blood stains and discovering latent blood stains.

Lasers or light sources for treating port-wine stains

DEFF Research Database (Denmark)

Faurschou, Annesofie; Olesen, Anne Braae; Leonardi-Bee, Jo

2011-01-01

Port-wine stains are birthmarks caused by malformations of blood vessels in the skin. Port-wine stains manifest themselves in infancy as a flat, red mark and do not regress spontaneously but may, if untreated, become darker and thicker in adult life. The profusion of various lasers and light...

An improved method for staining cell colonies in clonogenic assays.

Science.gov (United States)

Guda, Kishore; Natale, Leanna; Markowitz, Sanford D

2007-06-01

Clonogenic assay is a widely used experimental approach to test for the effects of drugs/genes on the growth and proliferative characteristics of cells in vitro. Accurate quantitation of treatment effects in clonogeneic assays depends on the ability to visualize and count cell colonies precisely. We report a novel method (referred as ETeB) for staining cell colonies grown on plastic and specially coated substrates like collagen. Using colon cancer cell lines grown on plastic and collagen, we compared the colony staining efficiencies of the widely used methylene blue, and Ethidium bromide (ETeB) stains. Results show that the ETeB protocol works well on plastic and is extremely effective for staining colonies on collagen when compared to methylene blue. The key features and advantages of ETeB technique are; (a) reduction in background for colonies grown on collagen and possibly other substrates, (b) the whole procedure takes less than a minute, (c) no post-stain washing step is required which eliminates colony losses for cell lines that are loosely adherent, (d) colony visualization and counting can be done immediately following the staining procedure using a standard UV illuminator and software, and (e) the method works across a wide variety of cell lines. The simplicity and robustness of this procedure should warrant its usage in both small and large-scale clonogenic experiments.

21 CFR 864.1850 - Dye and chemical solution stains.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Dye and chemical solution stains. 864.1850 Section 864.1850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Biological Stains § 864.1850 Dye and chemical...

Borax methylene blue: a spectroscopic and staining study.

Science.gov (United States)

Donaldson, P T; Russo, A; Reynolds, C; Lillie, R D

1978-07-01

Borax methylene blue is quite stable at room temperatures of 22-25 C. At 30 C polychroming is slow; during 50 days in a water bath at this temperature the absorption peak moves from 665 to 656 nm. At 35 C, the absorption peak reaches 660 nm in 7 days, 654 nm in 14. At 60 C polychroming is rapid, the absorption peak reaching 640-620 nm in 3 days. When the pH of the borax methylene blue solutions, normally about 9.0, is adjusted to pH 6.5, the absorption peak remains at 665 nm even when incubated at 60 C for extended periods. When used as a blood stain 0.4 ml borax methylene blue (1% methylene blue in 1% borax), 4 ml acetone, 2 ml borax-acid phosphate buffer to bring the solution to pH 6.5, and distilled water to make 40 ml, with 0.2 ml 1% eosin added just before using, an excellent Nocht-Giemsa type stain is achieved after 30 minutes staining. The material plasmodia P. falciparum, P. vivax, and P. berghei stain moderate blue with dark red chromatin and green to black pigment granules. The study confirms Malachowski's 1891 results and explains Gautier's 1896-98 failure to duplicate it.

A procedure for Alcian blue staining of mucins on polyvinylidene difluoride membranes.

Science.gov (United States)

Dong, Weijie; Matsuno, Yu-ki; Kameyama, Akihiko

2012-10-16

The isolation and characterization of mucins are critically important for obtaining insight into the molecular pathology of various diseases, including cancers and cystic fibrosis. Recently, we developed a novel membrane electrophoretic method, supported molecular matrix electrophoresis (SMME), which separates mucins on a polyvinylidene difluoride (PVDF) membrane impregnated with a hydrophilic polymer. Alcian blue staining is widely used to visualize mucopolysaccharides and acidic mucins on both blotted membranes and SMME membranes; however, this method cannot be used to stain mucins with a low acidic glycan content. Meanwhile, periodic acid-Schiff staining can selectively visualize glycoproteins, including mucins, but is incompatible with glycan analysis, which is indispensable for mucin characterizations. Here we describe a novel staining method, designated succinylation-Alcian blue staining, for visualizing mucins on a PVDF membrane. This method can visualize mucins regardless of the acidic residue content and shows a sensitivity 2-fold higher than that of Pro-Q Emerald 488, a fluorescent periodate Schiff-base stain. Furthermore, we demonstrate the compatibility of this novel staining procedure with glycan analysis using porcine gastric mucin as a model mucin.

Detection of chromogranins A and B in endocrine tissues with radioactive and biotinylated oligonucleotide probes

International Nuclear Information System (INIS)

Lloyd, R.V.; Jin, L.; Fields, K.

1990-01-01

We analyzed the distribution of chromogranins A and B in normal and neoplastic endocrine tissues with secretory granules using 35 S-labeled and biotin-labeled oligonucleotide probes by in situ hybridization (ISH). Both radioactive and nonradioactive probes detected messenger RNAs (mRNAs) in frozen and paraffin tissue sections. Endocrine tissues with variable immunoreactivities for chromogranin A protein, such as small-cell lung carcinomas, neuroblastomas, insulinomas, and parathyroid adenomas, expressed the mRNA for chromogranins A and B in most cells. Some technical problems with the biotinylated probes included nonspecific nuclear staining and endogenous alkaline phosphatase, which was not completely abolished by levamisole pretreatment. A differential distribution of chromogranins A and B was seen in pituitary prolactinomas, which expressed abundant chromogranin B but not chromogranin A mRNAs, and in parathyroid adenomas, which expressed abundant chromogranin A but only small amounts of chromogranin B mRNAs. These results indicate that ISH can be used to detect chromogranins A and B in endocrine tissues with radioactive and biotinylated oligonucleotide probes and that the mRNAs for chromogranin A and B are demonstrable in some tumors even when the chromogranin proteins cannot be detected by immunohistochemistry

Detection of chromogranins A and B in endocrine tissues with radioactive and biotinylated oligonucleotide probes

Energy Technology Data Exchange (ETDEWEB)

Lloyd, R.V.; Jin, L.; Fields, K. (Univ. of Michigan, Ann Arbor (USA))

1990-01-01

We analyzed the distribution of chromogranins A and B in normal and neoplastic endocrine tissues with secretory granules using {sup 35}S-labeled and biotin-labeled oligonucleotide probes by in situ hybridization (ISH). Both radioactive and nonradioactive probes detected messenger RNAs (mRNAs) in frozen and paraffin tissue sections. Endocrine tissues with variable immunoreactivities for chromogranin A protein, such as small-cell lung carcinomas, neuroblastomas, insulinomas, and parathyroid adenomas, expressed the mRNA for chromogranins A and B in most cells. Some technical problems with the biotinylated probes included nonspecific nuclear staining and endogenous alkaline phosphatase, which was not completely abolished by levamisole pretreatment. A differential distribution of chromogranins A and B was seen in pituitary prolactinomas, which expressed abundant chromogranin B but not chromogranin A mRNAs, and in parathyroid adenomas, which expressed abundant chromogranin A but only small amounts of chromogranin B mRNAs. These results indicate that ISH can be used to detect chromogranins A and B in endocrine tissues with radioactive and biotinylated oligonucleotide probes and that the mRNAs for chromogranin A and B are demonstrable in some tumors even when the chromogranin proteins cannot be detected by immunohistochemistry.

Improved Nissl method to stain formaldehyde or glutaraldehyde-fixed material.

Science.gov (United States)

Böck, P

1979-05-15

Nissl staining of paraffin sections from formaldehyde- or glutaraldehyde-fixed specimens is significantly intensified when sections are kept in a 50% (w/v) aqueous solution of potassium metabisulfite before being stained by a conventional Nissl method.

Technique and Feasibility of a Dual Staining Method for Estrogen Receptors and AgNORs

Directory of Open Access Journals (Sweden)

Lukas Günther

2000-01-01

Full Text Available A new staining method for dual demonstration of Estrogen receptors (ER and argyrophilc Nucleolus‐Organizer Regions (AgNORs was developed. To rule out possible reciprocal effects, serial slides of 10 invasive ductale breast cancers were stained with either the single staining method or the simultaneous ER/AgNOR‐staining method and investigated comparatively. By measuring the slides with the image analysis system AMBA, reciprocal effects could be excluded. It was proven that dual staining of both markers results in a reproducible and specific staining result. We concluded that it is justified to measure AgNORs in immunohistochemically stained cells.

Detection and Automated Scoring of Dicentric Chromosomes in Nonstimulated Lymphocyte Prematurely Condensed Chromosomes After Telomere and Centromere Staining

Energy Technology Data Exchange (ETDEWEB)

M' kacher, Radhia [Laboratoire de Radiobiologie et Oncologie, Commissariat à l' Energie Atomique, Fontenay-aux-Roses (France); El Maalouf, Elie [Laboratoire de Radiobiologie et Oncologie, Commissariat à l' Energie Atomique, Fontenay-aux-Roses (France); Laboratoire Modélisation Intelligence Processus Systèmes (MIPS)–Groupe TIIM3D, Université de Haute-Alsace, Mulhouse (France); Terzoudi, Georgia [Laboratory of Radiobiology & Biodosimetry, National Center for Scientific Research Demokritos, Athens (Greece); Ricoul, Michelle [Laboratoire de Radiobiologie et Oncologie, Commissariat à l' Energie Atomique, Fontenay-aux-Roses (France); Heidingsfelder, Leonhard [MetaSystems, Altlussheim (Germany); Karachristou, Ionna [Laboratory of Radiobiology & Biodosimetry, National Center for Scientific Research Demokritos, Athens (Greece); Laplagne, Eric [Pole Concept, Paris (France); Hempel, William M. [Laboratoire de Radiobiologie et Oncologie, Commissariat à l' Energie Atomique, Fontenay-aux-Roses (France); Colicchio, Bruno; Dieterlen, Alain [Laboratoire Modélisation Intelligence Processus Systèmes (MIPS)–Groupe TIIM3D, Université de Haute-Alsace, Mulhouse (France); Pantelias, Gabriel [Laboratory of Radiobiology & Biodosimetry, National Center for Scientific Research Demokritos, Athens (Greece); Sabatier, Laure, E-mail: laure.sabatier@cea.fr [Laboratoire de Radiobiologie et Oncologie, Commissariat à l' Energie Atomique, Fontenay-aux-Roses (France)

2015-03-01

Purpose: To combine telomere and centromere (TC) staining of premature chromosome condensation (PCC) fusions to identify dicentrics, centric rings, and acentric chromosomes, making possible the realization of a dose–response curve and automation of the process. Methods and Materials: Blood samples from healthy donors were exposed to {sup 60}Co irradiation at varying doses up to 8 Gy, followed by a repair period of 8 hours. Premature chromosome condensation fusions were carried out, and TC staining using peptide nucleic acid probes was performed. Chromosomal aberration (CA) scoring was carried out manually and automatically using PCC-TCScore software, developed in our laboratory. Results: We successfully optimized the hybridization conditions and image capture parameters, to increase the sensitivity and effectiveness of CA scoring. Dicentrics, centric rings, and acentric chromosomes were rapidly and accurately detected, leading to a linear-quadratic dose–response curve by manual scoring at up to 8 Gy. Using PCC-TCScore software for automatic scoring, we were able to detect 95% of dicentrics and centric rings. Conclusion: The introduction of TC staining to the PCC fusion technique has made possible the rapid scoring of unstable CAs, including dicentrics, with a level of accuracy and ease not previously possible. This new approach can be used for biological dosimetry in radiation emergency medicine, where the rapid and accurate detection of dicentrics is a high priority using automated scoring. Because there is no culture time, this new approach can also be used for the follow-up of patients treated by genotoxic therapy, creating the possibility to perform the estimation of induced chromosomal aberrations immediately after the blood draw.

Ocular melanoma metastatic to skin: the value of HMB-45 staining.

Science.gov (United States)

Schwartz, Robert A; Kist, Joseph M; Thomas, Isabelle; Fernández, Geover; Cruz, Manuel A; Koziorynska, Ewa I; Lambert, W Clark

2004-06-01

Cutaneous metastatic disease is an important finding that may represent the first sign of systemic cancer, or, if already known, that may change tumor staging and thus dramatically altered therapeutic plans. Although cutaneous metastases are relatively frequent in patients with cutaneous melanoma, they are less so from ocular melanoma. To demonstrate the value of HMB-45, staining in the detection of ocular melanoma metastatic to skin. The immunohistochemical stain HMB-45 a monoclonal antibody directed against intact human melanoma cells, was employed on a skin biopsy specimen from a cutaneous tumor. HMB-45 staining was positive in the atypical hyperchromatic cells of the deep dermis. HMB-45 may be of value in the detection of ocular melanoma metastatic to skin. Cutaneous metastatic disease is a somewhat common and extremely important diagnosis. Although cutaneous metastases from cutaneous melanoma are relatively frequent, those from ocular melanomas are less so. Use of histochemical staining, especially the HMB-45 stain, allows confirmation of the diagnosis.

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

NARCIS (Netherlands)

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

2011-01-01

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

[Intrapartum amnioinfusion in patients with meconium-stained amniotic fluid].

Science.gov (United States)

Engel, Karina; Samborska, Monika; Bilar, Marek; Sipak-Szmigiel, Olimpia; Ronin-Walknowska, Elzbieta

2008-09-01

The aim of the study was to evaluate the effect of intrapartum amnioinfusion in the presence of meconium stained amniotic fluid. 93 women with meconium-stained amniotic fluid were assigned to receive amnioinfusion or no amnioinfusion (128 women). The trials were evaluated for fetal distress syndrome, route of delivery, fetal acidemia, Apgar score at 1 and 5 min., meconium aspiration syndrome, postpartum endometritis and maternal hospital stays. Amnioinfusion in cases of meconium-stained fluid did not improve the number of fetal distress symptoms during fetal heart rate monitoring. Amnioinfusion was associated with a significant decrease of neonatal acidemia although it did not improve Apgar score. In our study amnioinfusion was not associated with reduction in the incidence of neonatal outcome and puerperial complications.

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

Science.gov (United States)

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

2012-08-01

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

Comparison of Gram and Kopeloff stains in the diagnosis of bacterial vaginosis in pregnancy.

Science.gov (United States)

Libman, Michael D; Kramer, Michael; Platt, Robert

2006-03-01

Bacterial vaginosis (BV) is commonly diagnosed by using the Nugent score, a semiquantitative scoring system to evaluate bacterial morphotypes on Gram stain of vaginal secretions. Some authors have suggested using the Kopeloff modification of the Gram stain. Asymptomatic BV in pregnancy has been associated with adverse outcomes. We performed both stains on simultaneously collected vaginal smears from 2652 women at 24-26 weeks of gestation. Gram staining gave significantly higher (more abnormal) Nugent scores than Kopeloff staining. Compared to the Kopeloff stain, the number of specimens graded as indeterminate or consistent with BV by Gram stain increased by 29% (469 versus 364, Pstaining was significantly better than Gram staining (agreement=74% versus 63%, intraclass correlation coefficient=0.87 versus 0.79, P<.05, 95% confidence intervals 0.85-0.89 and 0.75-0.82, respectively).

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

Science.gov (United States)

Cücer, N; Guler, N; Demirtas, H; Imamoğlu, N

2005-01-01

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

Improvement of malaria diagnostic system based on acridine orange staining.

Science.gov (United States)

Kimura, Masatsugu; Teramoto, Isao; Chan, Chim W; Idris, Zulkarnain Md; Kongere, James; Kagaya, Wataru; Kawamoto, Fumihiko; Asada, Ryoko; Isozumi, Rie; Kaneko, Akira

2018-02-07

Rapid diagnosis of malaria using acridine orange (AO) staining and a light microscope with a halogen lamp and interference filter was deployed in some malaria-endemic countries. However, it has not been widely adopted because: (1) the lamp was weak as an excitation light and the set-up did not work well under unstable power supply; and, (2) the staining of samples was frequently inconsistent. The halogen lamp was replaced by a low-cost, blue light-emitting diode (LED) lamp. Using a reformulated AO solution, the staining protocol was revised to make use of a concentration gradient instead of uniform staining. To evaluate this new AO diagnostic system, a pilot field study was conducted in the Lake Victoria basin in Kenya. Without staining failure, malaria infection status of about 100 samples was determined on-site per one microscopist per day, using the improved AO diagnostic system. The improved AO diagnosis had both higher overall sensitivity (46.1 vs 38.9%: p = 0.08) and specificity (99.0 vs 96.3%) than the Giemsa method (N = 1018), using PCR diagnosis as the standard. Consistent AO staining of thin blood films and rapid evaluation of malaria parasitaemia with the revised protocol produced superior results relative to the Giemsa method. This AO diagnostic system can be set up easily at low cost using an ordinary light microscope. It may supplement rapid diagnostic tests currently used in clinical settings in malaria-endemic countries, and may be considered as an inexpensive tool for case surveillance in malaria-eliminating countries.

[Automated analysis of bacterial preparations manufactured on automatic heat fixation and staining equipment].

Science.gov (United States)

2012-01-01

Heat fixation of preparations was made in the fixation bath designed by EMKO (Russia). Programmable "Emkosteiner" (EMKO, Russia) was used for trial staining. Reagents set Micko-GRAM-NITsF was applied for Gram's method of staining. It was demostrated that automatic smear fixation equipment and programmable staining ensure high-quality imaging (1% chromaticity variation) good enough for standardization of Gram's staining of microbial preparations.

Mechanisms of superficial micropunctate corneal staining with sodium fluorescein: the contribution of pooling.

Science.gov (United States)

Bandamwar, Kalika L; Garrett, Qian; Papas, Eric B

2012-04-01

To establish if sodium fluorescein (SFL) dye accumulation within intercellular spaces on the ocular surface contributes to the appearance of superficial punctate corneal staining. Thirteen subjects bilaterally wore PureVision™ lenses that had been pre-soaked in ReNu MultiPlus® multipurpose solution. After 1h of lens wear, corneal staining with SFL was assessed using a standard slit-lamp technique. Participants who presented with bilateral, corneal staining were selected for further evaluation. A randomly selected eye was rinsed with saline three times. Fellow eyes (control) received no rinsing. After each rinse, the appearance of SFL staining was recorded without any further instillation of the dye. To eliminate any confounding effects of staining due to residual fluorescein in the tear menisci, corneal staining was induced in freshly excised, isolated, rabbit eyes by topical administration of 0.001% PHMB and staining, rinsing and grading were performed as above. Nine out of 13 subjects presented with bilateral diffuse corneal staining (mean grade±SD: 2.4±0.7). The mean staining grades in test and control eyes respectively after each of the three rinses were (1) 2.41±0.41, 2.25±0.69 (p=0.9); (2) 2.34±0.79, 2.1±0.83 (p=0.8); and (3) 1.71±0.65, 1.60±0.79 (p=0.6) there was no significant reduction in staining with rinsing (p>0.05) and no difference was observed between test and control eyes at any sampling-point. Similar observations made in ex vivo rabbit eyes replicated these results. Pooling or accumulation of SFL solution within intercellular spaces does not appear to contribute to the appearance of superficial micropunctate corneal staining. Copyright © 2011 British Contact Lens Association. Published by Elsevier Ltd. All rights reserved.

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

Science.gov (United States)

Bautista, Pinky A; Yagi, Yukako

2012-05-01

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

Stain removal and whitening by baking soda dentifrice: A review of literature.

Science.gov (United States)

Li, Yiming

2017-11-01

Tooth discoloration may be caused by intrinsic or extrinsic stains or a combination of both. There are 2 major approaches to removing the stains, including the chemical mechanism using peroxides for tooth bleaching and the mechanical mechanism using abrasives in prophylactic pastes and dentifrices to remove stains, resulting in a whitening effect. Attempts have also been made to add a low concentration of peroxides to dentifrices to enhance their abrasive cleaning to remove tooth stains. This article provides a review of both in vitro and clinical studies on stain removal and whitening effect of dentifrices containing sodium bicarbonate (baking soda). In recent years, whitening dentifrices have become popular because of little additional effort for use, ease of availability, low cost, and accumulated evidence of clinical efficacy and safety in the literature. Advances in research and technology have led to innovative formulations of dentifrices using baking soda as the sole abrasive or a component of an abrasive system. Baking soda is biologically compatible with acid-buffering capacities, antibacterial at high concentrations, and has a relatively lower abrasivity. The evidence available in the literature indicates that baking soda-based dentifrices are effective and safe for tooth stain removal and consequently whitening. A number of clinical studies have also shown that baking soda-based dentifrices are more effective in stain removal and whitening than some non-baking soda-containing dentifrices with a higher abrasivity. So far, research efforts have mainly focused on stain removal and tooth-whitening efficacy and clinical safety of baking soda dentifrices used with manual toothbrushes, with only a few studies investigating their effects using powered toothbrushes, for which further research is encouraged. As part of a daily oral hygiene practice, baking soda-based dentifrice is a desirable, alternative or additional measure for tooth stain removal and whitening

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

International Nuclear Information System (INIS)

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

2015-01-01

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

The use of special stains in liver biopsy interpretation: Implications ...

African Journals Online (AJOL)

Materials and Methods: The formalin fixed paraffin embedded blocks of liver biopsies reported in two histopathology laboratories between 2008 and 2013 were retrieved. These were stained with H and E and the following standard special stains for liver tissue histology – Perl's Prussian blue, reticulin, Sirius red, Shikata ...

TANTANGAN DAN PELUANG JURUSAN TADRIS DI IAIN/STAIN

Directory of Open Access Journals (Sweden)

Mohammad Kosim

2009-01-01

Full Text Available A consequence of dualism in education policy, the implementatin of tadris in IAIN often faces many problems. This paper will describe the history and the development of tadris in IAIN/STAIN. The discussion begins with the origin of tadris in IAIN, problems araise in the implementation of tadris in IAIN/STAIN, including the alumni’s problem. In addition, this article also reveals several opportunities for tadris graduation to develop madrasah as purely religious school to public islamic school as the result of the paradigm change.

Utility of Acridine Orange staining for detection of bacteria from positive blood cultures.

Science.gov (United States)

Neeraja, M; Lakshmi, V; Padmasri, C; Padmaja, K

2017-08-01

The diagnostic performance of AO stain was evaluated for the detection of bacteria and or fungi from positive blood cultures. The sensitivity of Gram stain (GS) was 98.26% while Acridine Orange (AO) stain proved to be more sensitive (100%) with a Positive and Negative Predictive Value of 100% each. The specificity of both the stains was 100%. Overall agreement between the two stains was 98.23% (688/700). The organisms that were missed by GS and positive by AO were Candida species (Sutton, 2006) and Gram negative bacilli (GNB) (Sutton, 2006). Sensitivity of GS was 82.35% and AO was 100% among mixed cultures. Immediate reporting of the results of AO stain would have a significant impact on clinical management of patients with serious blood stream infections. Copyright © 2017 Elsevier B.V. All rights reserved.

Indirect immunofluorescence staining of Chlamydia trachomatis inclusions in microculture plates with monoclonal antibodies.

Science.gov (United States)

Zapata, M; Chernesky, M; Mahony, J

1984-06-01

Indirect immunofluorescence (IF) staining, using a monoclonal antibody, detected two- to fourfold more inclusions than did iodine staining. Of 274 clinical specimens, 53 (19.3%) were positive by IF on passage 1 as compared with 33 (12%) by iodine staining (P less than 0.005). IF-stained inclusions in McCoy cells in the bottom of microculture wells were readily viewed with a long-focal-length objective at a magnification of 250 X.

Indirect immunofluorescence staining of Chlamydia trachomatis inclusions in microculture plates with monoclonal antibodies.

OpenAIRE

Zapata, M; Chernesky, M; Mahony, J

1984-01-01

Indirect immunofluorescence (IF) staining, using a monoclonal antibody, detected two- to fourfold more inclusions than did iodine staining. Of 274 clinical specimens, 53 (19.3%) were positive by IF on passage 1 as compared with 33 (12%) by iodine staining (P less than 0.005). IF-stained inclusions in McCoy cells in the bottom of microculture wells were readily viewed with a long-focal-length objective at a magnification of 250 X.

Intracellular distribution of nontargeted quantum dots after natural uptake and microinjection

Science.gov (United States)

Damalakiene, Leona; Karabanovas, Vitalijus; Bagdonas, Saulius; Valius, Mindaugas; Rotomskis, Ricardas

2013-01-01

Background: The purpose of this study was to elucidate the mechanism of natural uptake of nonfunctionalized quantum dots in comparison with microinjected quantum dots by focusing on their time-dependent accumulation and intracellular localization in different cell lines. Methods: The accumulation dynamics of nontargeted CdSe/ZnS carboxyl-coated quantum dots (emission peak 625 nm) was analyzed in NIH3T3, MCF-7, and HepG2 cells by applying the methods of confocal and steady-state fluorescence spectroscopy. Intracellular colocalization of the quantum dots was investigated by staining with Lysotracker®. Results: The uptake of quantum dots into cells was dramatically reduced at a low temperature (4°C), indicating that the process is energy-dependent. The uptake kinetics and imaging of intracellular localization of quantum dots revealed three accumulation stages of carboxyl-coated quantum dots at 37°C, ie, a plateau stage, growth stage, and a saturation stage, which comprised four morphological phases: adherence to the cell membrane; formation of granulated clusters spread throughout the cytoplasm; localization of granulated clusters in the perinuclear region; and formation of multivesicular body-like structures and their redistribution in the cytoplasm. Diverse quantum dots containing intracellular vesicles in the range of approximately 0.5–8 μm in diameter were observed in the cytoplasm, but none were found in the nucleus. Vesicles containing quantum dots formed multivesicular body-like structures in NIH3T3 cells after 24 hours of incubation, which were Lysotracker-negative in serum-free medium and Lysotracker-positive in complete medium. The microinjected quantum dots remained uniformly distributed in the cytosol for at least 24 hours. Conclusion: Natural uptake of quantum dots in cells occurs through three accumulation stages via a mechanism requiring energy. The sharp contrast of the intracellular distribution after microinjection of quantum dots in comparison

Mobile Probing and Probes

DEFF Research Database (Denmark)

Duvaa, Uffe; Ørngreen, Rikke; Weinkouff Mathiasen, Anne-Gitte

2013-01-01

Mobile probing is a method, developed for learning about digital work situations, as an approach to discover new grounds. The method can be used when there is a need to know more about users and their work with certain tasks, but where users at the same time are distributed (in time and space......). Mobile probing was inspired by the cultural probe method, and was influenced by qualitative interview and inquiry approaches. The method has been used in two subsequent projects, involving school children (young adults at 15-17 years old) and employees (adults) in a consultancy company. Findings point...... to mobile probing being a flexible method for uncovering the unknowns, as a way of getting rich data to the analysis and design phases. On the other hand it is difficult to engage users to give in depth explanations, which seem easier in synchronous dialogs (whether online or face2face). The development...

Mobile Probing and Probes

DEFF Research Database (Denmark)

Duvaa, Uffe; Ørngreen, Rikke; Weinkouff, Anne-Gitte

2012-01-01

Mobile probing is a method, which has been developed for learning about digital work situations, as an approach to discover new grounds. The method can be used when there is a need to know more about users and their work with certain tasks, but where users at the same time are distributed (in time...... and space). Mobile probing was inspired by the cultural probe method, and was influenced by qualitative interview and inquiry approaches. The method has been used in two subsequent projects, involving school children (young adults at 15-17 years old) and employees (adults) in a consultancy company. Findings...... point to mobile probing being a flexible method for uncovering the unknowns, as a way of getting rich data to the analysis and design phases. On the other hand it is difficult to engage users to give in depth explanations, which seem easier in synchronous dialogs (whether online or face2face...

Photoacoustic Imaging of Port-Wine Stains

NARCIS (Netherlands)

Kolkman, R.G.M.; Mulder, M.J.; Mulder, Miranda J.; Glade, Conrad P.; Steenbergen, Wiendelt; van Leeuwen, Ton

2008-01-01

Background and Objective: To optimize laser therapy of port-wine stains (PWSs), information about the vasculature as well as lesion depth is valuable. In this study we investigated the use of photoacoustic imaging (PAI) to obtain this information. - Study Design/Materials and Methods: PAI uses

Identification criteria of the rare multi-flagellate Lophomonas blattarum: comparison of different staining techniques.

Science.gov (United States)

Alam-Eldin, Yosra Hussein; Abdulaziz, Amany Mamdouh

2015-09-01

Bronchopulmonary lophomoniasis (BPL) is an emerging disease of potential importance. BPL is presented by non-specific clinical picture and is usually accompanied by immunosuppression. Culture of Lophomonas blattarum is difficult and its molecular diagnosis has not yet been developed. Therefore, microscopic examination of respiratory samples, e.g., bronchoalveolar lavage (BAL) or sputum, is the mainstay of BPL diagnosis. Creola bodies and ciliocytophthoria are two forms of bronchial cells which occur in chest diseases with non-specific clinical picture like that of BPL. Both forms could be misrecognized as multi-flagellates because of their motile cilia in the wet mounts and due to shape variability of L. blattarum in stained smears. The aim of the study is to compare different staining techniques for visualizing L. blattarum to improve the recognition and diagnosis of BPL, to distinguish respiratory epithelial cells from L. blattarum and to decide which stain is recommended in suspected cases of BPL. BAL samples from patients which contain L. blattarum, creola bodies, and ciliocytophthoria were collected then wet mounts were examined. The BAL samples were also stained by Papanicolaou (PAP), Giemsa, hematoxylin and eosin (H & E), trichrome, Gram, and Diff-Quik (DQ) stains. The different staining techniques were compared regarding the stain quality. In wet mounts, the ciliary movement was coordinate and synchronous while the flagellar movement was wavy and leaded to active swimming of L. blattarum. In stained slides, bronchial cells were characterized by the presence of basal nucleus and the terminal bar from which the cilia arise. Trichrome was the best stain in demonstration of cellular details of L. blattarum. H & E, PAP, and Giemsa stains showed good quality of stains. Gram and DQ stains showed only pale hues of L. blattarum. We recommended adding Wheatley's trichrome staining to the differential diagnosis workup of cases of non-specific chest infections

Measurement of neuron soma size by fluorescent Nissl stain

OpenAIRE

sprotocols

2015-01-01

Authors: James Cronk, Noel Derecki & Jonathan Kipnis ### Abstract This protocol describes how to measure neuron soma size by fluorescent Nissl stain. Mice are sacrificed, and fixed by PFA perfusion. Brains are removed, and further PFA fixed, followed by sucrose cryoprotection. They are then snap frozen, sliced by cryostat, and stained with fluorescent Nissl as floating sections. Confocal microscopy is used to take images of neurons, and a computer graphics tablet is used to calculate ...

Evaluation of immunoperoxidase staining technique in the diagnosis of Acanthamoeba keratitis

Directory of Open Access Journals (Sweden)

Sharma Savitri

2001-01-01

Full Text Available Purpose: We describe a simple procedure of Immunoperoxidase (IP technique, using indigenously raised antibody, to screen corneal scrapings for Acanthamoeba cysts and trophozoites. This study sought to determine the utility of this test in the diagnosis of Acanthamoeba keratitis. Methods: A high titre polyclonal antibody against a local clinical isolate (axenic of Acanthamoeba species (trophozoite lysate antigen was raised in rabbits and used for standardization of IP technique for corneal scrapings. Twenty two smears of corneal scrapings, collected from patients showing Acanthamoeba cysts in corneal scrapings stained with calcofluorwhite (pool-1 and patients showing no cysts in similar scrapings (pool-2, were coded and stained by IP technique by a masked technician. All 22 patients had also been tested for bacteria, fungus, and Acanthamoeba in their corneal scrapings by smears and cultures. IP stained smears were examined for organisms including cysts and trophozoites of Acanthamoeba and background staining by two observers masked to the results of other smears and cultures. The validity of the IP test in detection of Acanthamoeba cysts and trophozoites was measured by sensitivity, specificity, positive predictive value and negative predictive value in comparison (McNemar test for paired comparison with calcofluor white staining and culture. Results: Based on the readings of observer 1 and compared to calcofluor white staining, the IP test had a sensitivity of 100%, a specificity of 94%, positive predictive value of 80% and negative predictive value of 100%. When compared to culture, the values were 83%, 100%, 100% and 94% respectively. Trophozoites missed in calcofluor white stained smears, were detected in 2 out of 6 cases of culture-positive Acanthamoeba keratitis. The Kappa coefficient of interobserver agreement was determined as fair (30.4%. Conclusion: The immunoperoxidase technique is a simple and useful test in the diagnosis of

Hierarchical patch-based co-registration of differently stained histopathology slides

Science.gov (United States)

Yigitsoy, Mehmet; Schmidt, Günter

2017-03-01

Over the past decades, digital pathology has emerged as an alternative way of looking at the tissue at subcellular level. It enables multiplexed analysis of different cell types at micron level. Information about cell types can be extracted by staining sections of a tissue block using different markers. However, robust fusion of structural and functional information from different stains is necessary for reproducible multiplexed analysis. Such a fusion can be obtained via image co-registration by establishing spatial correspondences between tissue sections. Spatial correspondences can then be used to transfer various statistics about cell types between sections. However, the multi-modal nature of images and sparse distribution of interesting cell types pose several challenges for the registration of differently stained tissue sections. In this work, we propose a co-registration framework that efficiently addresses such challenges. We present a hierarchical patch-based registration of intensity normalized tissue sections. Preliminary experiments demonstrate the potential of the proposed technique for the fusion of multi-modal information from differently stained digital histopathology sections.

Photoacoustic imaging of port-wine stains

NARCIS (Netherlands)

Kolkman, Roy G. M.; Mulder, Miranda J.; Glade, Conrad P.; Steenbergen, Wiendelt; van Leeuwen, Ton G.

2008-01-01

BACKGROUND AND OBJECTIVE: To optimize laser therapy of port-wine stains (PWSs), information about the vasculature as well as lesion depth is valuable. In this study we investigated the use of photoacoustic imaging (PAI) to obtain this information. STUDY DESIGN/MATERIALS AND METHODS: PAI uses pulsed

Colour stability, staining and roughness of silorane after prolonged chemical challenges

DEFF Research Database (Denmark)

Benetti, Ana Raquel; Ribeiro de Jesus, Vivian Cristiane Bueno; Martinelli, Natan Luiz

2013-01-01

methacrylate or silorane composites. Specimens were individually stored at 37°C in 0.02 N citric acid, 0.02 N phosphoric acid, 75% ethanol or distilled water for 7, 14, 21 and 180 days, when new measurements were performed. A staining test was performed after the chemical challenge by immersion in coffee...... considered acceptable (although significantly different) after immersion in water, citric acid, phosphoric acid or ethanol, but were unacceptable for the silorane composite immersed in ethanol for 180 days. The methacrylate-based resins stored in ethanol were significantly more stained by coffee than those...... stored in other media. The silorane composite demonstrated no staining, but increased roughness, when compared to the methacrylate-based resins. CONCLUSIONS: No effect of the immersion solution was noticed on roughness of the investigated materials. Ethanol influenced colour stability and staining...

Automated robust registration of grossly misregistered whole-slide images with varying stains

Science.gov (United States)

Litjens, G.; Safferling, K.; Grabe, N.

2016-03-01

Cancer diagnosis and pharmaceutical research increasingly depend on the accurate quantification of cancer biomarkers. Identification of biomarkers is usually performed through immunohistochemical staining of cancer sections on glass slides. However, combination of multiple biomarkers from a wide variety of immunohistochemically stained slides is a tedious process in traditional histopathology due to the switching of glass slides and re-identification of regions of interest by pathologists. Digital pathology now allows us to apply image registration algorithms to digitized whole-slides to align the differing immunohistochemical stains automatically. However, registration algorithms need to be robust to changes in color due to differing stains and severe changes in tissue content between slides. In this work we developed a robust registration methodology to allow for fast coarse alignment of multiple immunohistochemical stains to the base hematyoxylin and eosin stained image. We applied HSD color model conversion to obtain a less stain color dependent representation of the whole-slide images. Subsequently, optical density thresholding and connected component analysis were used to identify the relevant regions for registration. Template matching using normalized mutual information was applied to provide initial translation and rotation parameters, after which a cost function-driven affine registration was performed. The algorithm was validated using 40 slides from 10 prostate cancer patients, with landmark registration error as a metric. Median landmark registration error was around 180 microns, which indicates performance is adequate for practical application. None of the registrations failed, indicating the robustness of the algorithm.

News from the Biological Stain Commission no. 12

DEFF Research Database (Denmark)

Lyon, H O

2012-01-01

In this 12(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 meetings of ISO/TC 212/WG 1 Quality and competence in the medical laboratory and ISO....../TC 212/WG 3 In vitro diagnostic products both held on 2 - 3 June 2010, plus information on the second plenary meeting of ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on 4 June 2010. All meetings took place in Seoul, Republic of Korea. Finally, information is provided...

[Comparison of four different staining methods for ear cytology of dogs with otitis externa].

Science.gov (United States)

Bouassiba, C; Osthold, W; Mueller, R S

2013-01-01

Cytological examination is crucial for the diagnosis and classification of canine otitis externa. Staining should reveal micro-organisms as perpetuating factors of otitis externa. The aim of the study was to compare four different staining methods (Diff-Quik®, Diff-Quik® after dipping in acetone, Gram Quick stain® and a commercial rapid stain for otitis externa) for ear cytology of dogs with otitis externa and to investigate the agreement of cytology and culture. In a study evaluating dogs with otitis externa, five ear swabs (one for culture and four for cytology) were taken from the horizontal part of the external auditory canal of 224 affected ears and compared semi-quantitatively. Diff-Quik® with and without prior dipping in acetone as well as the Gram Quick stain® displayed a high degree of agreement in the detection of micro-organisms (cocci p = 0.2366; rods p = 0.4832; yeasts p = 0.1574), while the commercial otitis rapid stain revealed significantly less micro-organisms (p stains corresponded to the culture results by >  70%; the agreement was lower with the commercial otitis rapid stain. The quickest and easiest method was staining with Diff-Quik®. Diff-Quik® with or without prior dipping in acetone and the Gram Quick stain® had a high agreement in the detection of microorganisms and can thus be considered nearly equivalent for the diagnosis of otitis externa infectiosa. The commercial otitis rapid stain is less reliable. Based on this study Diff-Quik® can be recommended for the routine cytology of ear swabs. Additionally, a culture may be indicated and must be interpreted in the context of the cytology.

A low molecular weight zinc{sup 2+}-dipicolylamine-based probe detects apoptosis during tumour treatment better than an annexin V-based probe

Energy Technology Data Exchange (ETDEWEB)

Palmowski, Karin [RWTH-Aachen University, Department of Experimental Molecular Imaging, Aachen (Germany); University of Heidelberg, Department of Pneumology, Thoraxklinik Heidelberg, Heidelberg (Germany); Rix, Anne; Lederle, Wiltrud; Kiessling, Fabian [RWTH-Aachen University, Department of Experimental Molecular Imaging, Aachen (Germany); Behrendt, Florian F. [RWTH-Aachen University, Department of Nuclear Medicine, Aachen (Germany); Mottaghy, Felix M. [RWTH-Aachen University, Department of Nuclear Medicine, Aachen (Germany); Maastricht University Medical Center, Department of Nuclear Medicine, Maastricht (Netherlands); Gray, Brian D. [Molecular Targeting Technologies, Inc., West Chester, PA (United States); Pak, Koon Y. [University Medical Center Heidelberg, Academic Radiology Baden-Baden, Diagnostic and Interventional Radiology, Heidelberg (Germany); Palmowski, Moritz [RWTH-Aachen University, Department of Experimental Molecular Imaging, Aachen (Germany); RWTH-Aachen University, Department of Nuclear Medicine, Aachen (Germany); University Medical Center Heidelberg, Academic Radiology Baden-Baden, Diagnostic and Interventional Radiology, Heidelberg (Germany)

2014-02-15

Molecular imaging of apoptosis is frequently discussed for monitoring cancer therapies. Here, we compare the low molecular weight phosphatidylserine-targeting ligand zinc{sup 2+}-dipicolylamine (Zn{sup 2+}-DPA) with the established but reasonably larger protein annexin V. Molecular apoptosis imaging with the fluorescently labelled probes annexin V (750 nm, 36 kDa) and Zn{sup 2+}-DPA (794 nm, 1.84 kDa) was performed in tumour-bearing mice (A431). Three animal groups were investigated: untreated controls and treated tumours after 1 or 4 days of anti-angiogenic therapy (SU11248). Additionally, μPET with {sup 18} F-FDG was performed. Imaging data were displayed as tumour-to-muscle ratio (TMR) and validated by quantitative immunohistochemistry. Compared with untreated control tumours, TUNEL staining indicated significant apoptosis after 1 day (P < 0.05) and 4 days (P < 0.01) of treatment. Concordantly, Zn{sup 2+}-DPA uptake increased significantly after 1 day (P < 0.05) and 4 days (P < 0.01). Surprisingly, annexin V failed to detect significant differences between control and treated animals. Contrary to the increasing uptake of Zn{sup 2+}-DPA, {sup 18} F-FDG tumour uptake decreased significantly at days 1 (P < 0.05) and 4 (P < 0.01). Increase in apoptosis during anti-angiogenic therapy was detected significantly better with the low molecular weight probe Zn{sup 2+}-DPA than with the annexin V-based probe. Additionally, significant treatment effects were detectable as early using Zn{sup 2+}-DPA as with measurements of the glucose metabolism using {sup 18} F-FDG. (orig.)

[Improvement of Phi bodies stain and its clinical significance].

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Gong, Xu-Bo; Lu, Xing-Guo; Yan, Li-Juan; Xiao, Xi-Bin; Wu, Dong; Xu, Gen-Bo; Zhang, Xiao-Hong; Zhao, Xiao-Ying

2009-02-01

The aim of this study was to improve the dyeing method of hydroperoxidase (HPO), to analyze the morphologic features of Phi bodies and to evaluate the clinical application of this method. 128 bone marrow or peripheral blood smears from patients with myeloid and lymphoid malignancies were stained by improved HPO staining. The Phi bodies were observed with detection rate of Phi bodies in different leukemias. 69 acute myeloid leukemia (AML) specimens were chosen randomly, the positive rate and the number of Phi bodies between the improved HPO and POX stain based on the same substrate of 3, 3'diaminobenzidine were compared. The results showed that the shape of bundle-like Phi bodies was variable, long or short. while the nubbly Phi bodies often presented oval and smooth. Club-like Phi bodies were found in M(3). The detection rates of bundle-like Phi bodies in AML M(1)-M(5) were 42.9% (6/14), 83.3% (15/18), 92.0% (23/25), 52.3% (11/21), 33.3% (5/15) respectively, and those of nubbly Phi bodies were 28.6% (4/14), 66.7% (12/18), 11.1% (3/25), 33.3% (7/21), 20.0% (3/15) respectively. The detection rate of bundle-like Phi bodies in M(3) was significantly higher than that in (M(1) + M(2)) or (M(4) + M(5)) groups. The detection rate of nubbly Phi bodies in (M(1) + M(2)) group was higher than that in M(3) group. In conclusion, after improvement of staining method, the HPO stain becomes simple, the detection rate of Phi bodies is higher than that by the previous method, the positive granules are more obvious, and the results become stable. This improved method plays an important role in differentiating AML from ALL, subtyping AML, and evaluating the therapeutic results.

Systematic investigation of drip stains on apparel fabrics: The effects of prior-laundering, fibre content and fabric structure on final stain appearance.

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de Castro, Therese C; Taylor, Michael C; Kieser, Jules A; Carr, Debra J; Duncan, W

2015-05-01

Bloodstain pattern analysis is the investigation of blood deposited at crime scenes and the interpretation of that pattern. The surface that the blood gets deposited onto could distort the appearance of the bloodstain. The interaction of blood and apparel fabrics is in its infancy, but the interaction of liquids and apparel fabrics has been well documented and investigated in the field of textile science (e.g. the processes of wetting and wicking of fluids on fibres, yarns and fabrics). A systematic study on the final appearance of drip stains on torso apparel fabrics (100% cotton plain woven, 100% polyester plain woven, blend of polyester and cotton plain woven and 100% cotton single jersey knit) that had been laundered for six, 26 and 52 cycles prior to testing was investigated in the paper. The relationship between drop velocity (1.66±0.50m/s, 4.07±0.03m/s, 5.34±0.18m/s) and the stain characteristics (parent stain area, axes 1 and 2 and number of satellite stains) for each fabric was examined using analysis of variance. The experimental design and effect of storing blood were investigated on a reference sample, which indicated that the day (up to five days) at which the drops were generated did not affect the bloodstain. The effect of prior-laundering (six, 26 and 52 laundering cycles), fibre content (cotton vs. polyester vs. blend) and fabric structure (plain woven vs. single jersey knit) on the final appearance of the bloodstain were investigated. Distortion in the bloodstains produced on non-laundered fabrics indicated the importance of laundering fabrics to remove finishing treatments before conducting bloodstain experiments. For laundered fabrics, both the cotton fabrics and the blend had a circular to oval stain appearance, while the polyester fabric had a circular appearance with evidence of spread along the warp and weft yarns, which resulted in square-like stains at the lowest drop velocity. A significant (pfibre content (pfibres/yarns, while for the

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

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Wahlsten, Douglas; Colbourne, Frederick; Pleus, Richard

2003-03-15

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

NIST mixed stain study 3: signal intensity balance in commercial short tandem repeat multiplexes.

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Duewer, David L; Kline, Margaret C; Redman, Janette W; Butler, John M

2004-12-01

Short-tandem repeat (STR) allelic intensities were collected from more than 60 forensic laboratories for a suite of seven samples as part of the National Institute of Standards and Technology-coordinated 2001 Mixed Stain Study 3 (MSS3). These interlaboratory challenge data illuminate the relative importance of intrinsic and user-determined factors affecting the locus-to-locus balance of signal intensities for currently used STR multiplexes. To varying degrees, seven of the eight commercially produced multiplexes used by MSS3 participants displayed very similar patterns of intensity differences among the different loci probed by the multiplexes for all samples, in the hands of multiple analysts, with a variety of supplies and instruments. These systematic differences reflect intrinsic properties of the individual multiplexes, not user-controllable measurement practices. To the extent that quality systems specify minimum and maximum absolute intensities for data acceptability and data interpretation schema require among-locus balance, these intrinsic intensity differences may decrease the utility of multiplex results and surely increase the cost of analysis.

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

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

2014-01-01

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

Smart near-infrared fluorescence probes with donor-acceptor structure for in vivo detection of β-amyloid deposits.

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Cui, Mengchao; Ono, Masahiro; Watanabe, Hiroyuki; Kimura, Hiroyuki; Liu, Boli; Saji, Hideo

2014-03-05

The deposition of β-amyloid (Aβ) plaques in the parenchymal and cortical brain is accepted as the main pathological hallmark of Alzheimer's disease (AD); however, early detection of AD still presents a challenge. With the assistance of molecular imaging techniques, imaging agents specifically targeting Aβ plaques in the brain may lead to the early diagnosis of AD. Herein, we report the design, synthesis, and evaluation of a series of smart near-infrared fluorescence (NIRF) imaging probes with donor-acceptor architecture bridged by a conjugated π-electron chain for Aβ plaques. The chemical structure of these NIRF probes is completely different from Congo Red and Thioflavin-T. Probes with a longer conjugated π system (carbon-carbon double bond) displayed maximum emission in PBS (>650 nm), which falls in the best range for NIRF probes. These probes were proved to have affinity to Aβ plaques in fluorescent staining of brain sections from an AD patient and double transgenic mice, as well as in an in vitro binding assay using Aβ(1-42) aggregates. One probe with high affinity (K(i) = 37 nM, K(d) = 27 nM) was selected for in vivo imaging. It can penetrate the blood-brain barrier of nude mice efficiently and is quickly washed out of the normal brain. Moreover, after intravenous injection of this probe, 22-month-old APPswe/PSEN1 mice exhibited a higher relative signal than control mice over the same period of time, and ex vivo fluorescent observations confirmed the existence of Aβ plaques. In summary, this probe meets most of the requirements for a NIRF contrast agent for the detection of Aβ plaques both in vitro and in vivo.

Investigation Of The Color Changing Properties Of Wood Stain Derived From Pinar Leaves

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Abdi Atılgan

2011-11-01

Full Text Available This study was designed to develop an environmentally friendly wood stain derived pinar (Quercus aucheri leaves and determine the color stability of this stain when exposed to UV light irradiation. Wood stains derived from pinar leaves were prepared from aqueous solution with %3 iron (FeSO4.7H2O , % 5 alum ((KAl(SO42.12H2O, and % 10 vinegar mordant mixtures. Scots pine (Pinus sylvestris L., Turkish oriental beech (Fagus orientalis Lipsky and oak (Quercus petraea L. wood specimens were used as staining substrates. After treatment with the stain, the wood panels were exposed to UV light irradiation for periods of 100, 200, and 300 hours and determinated the total color changes was according to ISO 2470 standards. Results showed that wood stain derived from pinar extract provided some color stability after UV irradiation. According to results, Scots pine specimens treated with the pinar extract + iron mixture provided the smallest total color changes. Meanwhile the highest total color change provided on the Scots pine treated with pinar extract+alum mixture.

MEGARA Optics: stain removal in PBM2Y prisms

International Nuclear Information System (INIS)

Aguirre-Aguirre, D; Izazaga-Pérez, R; Carrasco, E; Villalobos-Mendoza, B; De Paz, A Gil; Gallego, J; Iglesias, J

2017-01-01

MEGARA is the new integral-field and multi-object optical spectrograph for the GTC. For medium and high resolution, the dispersive elements are volume phase holographic gratings, sandwiched between two flat windows and two prisms of high optical precision. The prisms are made of Ohara PBM2Y optical glass. After the prisms polishing process, some stains appeared on the surfaces. For this, in this work is shown the comparative study of five different products (muriatic acid, paint remover, sodium hydroxide, aqua regia and rare earth liquid polish) used for trying to eliminate the stains of the HR MEGARA prisms. It was found that by polishing with the hands the affected area, and using a towel like a kind of pad, and polish during five minutes using rare earth, the stains disappear completely affecting only a 5% the rms of the surface quality. Not so the use of the other products that did not show any apparent result. (paper)

Neo-Timm staining in the thalamus of chronically epileptic rats

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Hamani C.

2005-01-01

Full Text Available The thalamus is an important modulator of seizures and is severely affected in cholinergic models of epilepsy. In the present study, chronically epileptic rats had their brains processed for neo-Timm and acetylcholinesterase two months after the induction of status epilepticus with pilocarpine. Both controls and pilocarpine-treated animals presented neo-Timm staining in the anterodorsal nucleus, laterodorsal nucleus, reticular nucleus, most intralaminar nuclei, nucleus reuniens, and rhomboid nucleus of the thalamus, as well as in the zona incerta. The intensity of neo-Timm staining was similar in control and pilocarpine-treated rats, except for the nucleus reuniens and the rhomboid nucleus, which had a lower intensity of staining in the epileptic group. In animal models of temporal lobe epilepsy, zinc seems to modulate glutamate release and to decrease seizure activity. In this context, a reduction of neo-Timm-stained terminals in the midline thalamus could ultimately result in an increased excitatory activity, not only within its related nuclei, but also in anatomical structures that receive their efferent connections. This might contribute to the pathological substrate observed in chronic pilocarpine-treated epileptic animals.

An improved silver staining procedure for schizodeme analysis in polyacrylamide gradient gels

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Antonio M. Gonçalves

1990-03-01

Full Text Available A simple protocol is described for the silver staining of polyacrylamide gradient gels used for the separation of restriction fragments of kinetoplast DNA [schizodeme analysis of trypanosomatids (Morel et al., 1980]. The method overcomes the problems of non-uniform staining and strong background color which are frequently encountered when conventional protocols for silver staining of linear gels. The method described has proven to be of general applicability for DNA, RNA and protein separations in gradient gels.

Efficacy of evaluation of rooster sperm morphology using different staining methods.

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Lukaszewicz, E; Jerysz, A; Partyka, A; Siudzińska, A

2008-12-01

This work focused on inexpensive methods of evaluation fowl sperm morphology, based on eosin-nigrosin smears, which can determine disorders in spermatogenesis and can be recommended for evaluating the fertilising potency and selecting males in flocks reproduced by artificial insemination. Four fowl breeds (Black Minorca, Italian Partridge, Forwerk and Greenleg Partridge) were used to determine the efficacy of sperm morphology evaluation using four eosin-nigrosin staining methods (according to Blom, Bakst and Cecil, Morisson, Jaśkowski) and three examiners of different experience (high, medium, novice). There were significant (P< or = 0.01) differences in sperm morphology between Blom's staining method and those of Bakst and Cecil, Morisson or Jaśkowski, irrespective of fowl breed and examiners experience. Blom stain caused sperm head swelling and showed a drastic reduction in the proportion of live spermatozoa with normal morphology. The staining method had a greater influence on sperm morphology evaluation than the experience of the examiners.

A combined Bodian-Nissl stain for improved network analysis in neuronal cell culture.

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Hightower, M; Gross, G W

1985-11-01

Bodian and Nissl procedures were combined to stain dissociated mouse spinal cord cells cultured on coverslips. The Bodian technique stains fine neuronal processes in great detail as well as an intracellular fibrillar network concentrated around the nucleus and in proximal neurites. The Nissl stain clearly delimits neuronal cytoplasm in somata and in large dendrites. A combination of these techniques allows the simultaneous depiction of neuronal perikarya and all afferent and efferent processes. Costaining with little background staining by either procedure suggests high specificity for neurons. This procedure could be exploited for routine network analysis of cultured neurons.

Immunohistochemical positive stained p53 protein in bladder transitional cell carcinoma

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Halimi Monireh

2009-04-01

Full Text Available Background: Molecular genetics and immunopathologic analysis of bladder cancer have shown some abnormalities in a number of genes and proteins that have been implicated in the development and progression of such tumors, mainly in the p53 pathway. Aims: To investigate the rate of positively stained p53 protein in patients with urothelial papillary carcinoma of the bladder (UCB by immunohistochemistry and its relationship with tumor grade, gender and age of the patients. Settings and Design: During the present cross-sectional study, 100 paraffin-embedded specimens of UCB, which were provided from biopsies of the bladder by transurethral access, were immunohistochemically stained and studied for p53 protein from May 2006 to May 2007 in our referral center pathology laboratory. Materials and Methods: First, 4 µm slices of paraffin sections were provided and then stained by the avidin-biotin peroxidase method. The rate of positively stained p53 protein (defined as positive nuclear staining in over 10% of the cells was assessed. This rate was also estimated and compared between grades, genders and age-related groups (< 70 years, ≥70 years. Statistical Analysis: The χ2 , Fisher′s exact test and Mann-Whitney U test were used for comparing. Results: The overall rate of positively stained specimens was 11% for nuclear p53 protein. This rate was significantly higher in females (10/29 vs. 1/71; P < 0.001; odds ratio [OR]: 0.23; 95% confidence interval [CI]: 4.43-306.08, patients with 70 or older than 70 years (8/42 vs. 3/58; P = 0.04; OR: 0.55; 95% CI: 1.07-17.39 and in high-grade tumors (10/58 vs. 1/42; P = 0.02; OR: 0.59; 95% CI: 0.01-0.95. Conclusions: The rate of positively stained p53 protein for UCB was lower in our population. This rate was also higher in females, patients with 70 or older than 70 years and high grade of UCB.

Effectiveness of clean-up procedures on stain susceptibility of different orthodontic adhesives

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Swati Pundlik Mane

2014-01-01

Conclusion: Chemical-cure adhesive showed higher stain susceptibility than light-cure adhesive in all clean-up procedures. Both adhesives would show less stain susceptibility with polishing step with rubber cup and pumice.

Staining of proteins in gels with Coomassie G-250 without organic solvent and acetic acid.

Science.gov (United States)

Lawrence, Ann-Marie; Besir, H Uuml Seyin

2009-08-14

In classical protein staining protocols using Coomassie Brilliant Blue (CBB), solutions with high contents of toxic and flammable organic solvents (Methanol, Ethanol or 2-Propanol) and acetic acid are used for fixation, staining and destaining of proteins in a gel after SDS-PAGE. To speed up the procedure, heating the staining solution in the microwave oven for a short time is frequently used. This usually results in evaporation of toxic or hazardous Methanol, Ethanol or 2-Propanol and a strong smell of acetic acid in the lab which should be avoided due to safety considerations. In a protocol originally published in two patent applications by E.M. Wondrak (US2001046709 (A1), US6319720 (B1)), an alternative composition of the staining solution is described in which no organic solvent or acid is used. The CBB is dissolved in bidistilled water (60-80 mg of CBB G-250 per liter) and 35 mM HCl is added as the only other compound in the staining solution. The CBB staining of the gel is done after SDS-PAGE and thorough washing of the gel in bidistilled water. By heating the gel during the washing and staining steps, the process can be finished faster and no toxic or hazardous compounds are evaporating. The staining of proteins occurs already within 1 minute after heating the gel in staining solution and is fully developed after 15-30 min with a slightly blue background that is destained completely by prolonged washing of the stained gel in bidistilled water, without affecting the stained protein bands.

Efficacy of baking soda-containing chewing gum in removing natural tooth stain.

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Mankodi, S M; Conforti, N; Berkowitz, H

2001-07-01

A 14-week, double-blind, randomized clinical trial was conducted with 126 healthy volunteers to compare the efficacy of twice-daily use of 3 baking soda-containing chewing gums in removing natural tooth stain when used in conjunction with a program of regular oral hygiene. All 3 chewing gums significantly reduced extrinsic stain (P Baking Soda Gum (AHDC) reduced dental stain by 70.8%, compared to reductions of 71.9% and 65.3%, after use of 2 experimental gum formulations. Whitened appearance improved by 1.73 shade tabs using AHDC gum, and up to 2.49 shade tabs with the experimental formulations. These results suggest that the use of baking soda-containing gum after meals, in conjunction with good oral hygiene, can improve both extrinsic dental staining and the whitened appearance of teeth.

Evaluation Method of Accumulated Cooking Oil Stains in Room

OpenAIRE

五十嵐, 由利子; 中村, 和吉; 萬羽, 郁子; Igarashi, Yuriko; Nakamura, Kazuyoshi; Banba, Ikuko

2005-01-01

The diffusion of the oil mist while cooking affects the entire room, leaving stains on the ceiling and walls. The validity of measuring the color difference of stains was examined by installing teflon plates in the kitchen and the adjacent space with a view to assessing the oil diffusion. Four houses were designated for the examination. In each house, four teflon plates (20×40mm) were installed on the ceiling and walls of the kitchen and the space adjacent to it. Then, one plate was removed a...

Improving Gram stain proficiency in hospital and satellite laboratories that do not have microbiology.

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Guarner, Jeannette; Street, Cassandra; Matlock, Margaret; Cole, Lisa; Brierre, Francoise

2017-03-01

Consolidation of laboratories has left many hospitals and satellite laboratories with minimal microbiologic testing. In many hospitals and satellite laboratories, Gram stains on primary specimens are still performed despite difficultly in maintaining proficiency. To maintain Gram stain proficiency at a community 450-bed hospital with an active emergency room we designed bimonthly challenges that require reporting Gram staining and morphology of different organisms. The challenges consist of five specimens prepared by the reference microbiology laboratory from cultures and primary specimens. Twenty to 23 medical laboratory scientists participate reading the challenges. Results from the challenges are discussed with each medical laboratory scientists. In addition, printed images from the challenges are presented at huddle to add microbiology knowledge. On the first three challenges, Gram staining was read correctly in 71%-77% of the time while morphology 53%-66%. In the last six challenges correct answers for Gram stain were 77%-99% while morphology 73%-96%. We observed statistically significant improvement when reading Gram stains by providing frequent challenges to medical laboratory scientists. The clinical importance of Gram stain results is emphasized during huddle presentations increasing knowledge and motivation to perform the test for patients.

Amazonian açai and food dyes for staining arbuscular- micorrhizal fungi

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Aline Lourdes Martins Silva

2015-12-01

Full Text Available Arbuscular mycorrhizae microscopy requires differential staining of typical structures. Dyes employed, such as trypan blue, pose risks to health and environment. Alternative dyes such as pen ink and aniline have variable coloring efficiency. In this work, Brachiaria decumbens roots, discolored with caustic soda (NaOH, were stained with açai, annatto, saffron, trypan blue and pen inks. There were significant differences among dyes regarding stained mycorrhizal structures and pictures quality. Acai was considered the best alternative dye, with similar results to trypan blue.

Nanoparticle-based luminescent probes for intracellular sensing and imaging of pH.

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Schäferling, Michael

2016-05-01

Fluorescence imaging microscopy is an essential tool in biomedical research. Meanwhile, various fluorescent probes are available for the staining of cells, cell membranes, and organelles. Though, to monitor intracellular processes and dysfunctions, probes that respond to ubiquitous chemical parameters determining the cellular function such as pH, pO2 , and Ca(2+) are required. This review is focused on the progress in the design, fabrication, and application of photoluminescent nanoprobes for sensing and imaging of pH in living cells. The advantages of using nanoprobes carrying fluorescent pH indicators compared to single molecule probes are discussed as well as their limitations due to the mostly lysosomal uptake by cells. Particular attention is paid to ratiometric dual wavelength nanosensors that enable intrinsic referenced measurements. Referencing and proper calibration procedures are basic prerequisites to carry out reliable quantitative pH determinations in complex samples such as living cells. A variety of examples will be presented that highlight the diverseness of nanocarrier materials (polymers, micelles, silica, quantum dots, carbon dots, gold, photon upconversion nanocrystals, or bacteriophages), fluorescent pH indicators for the weak acidic range, and referenced sensing mechanisms, that have been applied intracellularly up to now. WIREs Nanomed Nanobiotechnol 2016, 8:378-413. doi: 10.1002/wnan.1366 For further resources related to this article, please visit the WIREs website. © 2015 Wiley Periodicals, Inc.

Study of stained glass window using PIXE-PIGE

International Nuclear Information System (INIS)

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

2005-01-01

We had the opportunity to study a large panel (100x80cm) containing more than 40 stained glass pieces. Among them several come from restorations having taken place at different periods. The study of this rather complex arrangement has been processed by stages:- the elemental composition of 16 zones were determined: several differences were identified and among them the Na/K ratio which allowed to set three groups of glass type; - the measurement of the Na concentrations by the two techniques give information in bulk (PIGE) and at the near surface (PIXE); the values defined by the (C PIGE -C PIXE) )/C PIGE plotted in function of the historical estimation of the age of the stained glass pieces (original and restored) indicate a real correlation between the two variables; - the red-colored pieces were specially investigated in order to determine which coloration technique was employed (bulk coloration, superficial staining, multilayered flashing, etc.); - the corrosion was investigated by scanning two different worsened zones with a 0.5mm diameter beam spot. This study shows the possibilities of the PIGE-PIXE association, but also points out some weaknesses, which have to be solved by other techniques; unfortunately, in that case, the non-destructive aspect could be lost

Influence of probe motion on laser probe temperature in circulating blood.

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Hehrlein, C; Splinter, R; Littmann, L; Tuntelder, J R; Tatsis, G P; Svenson, R H

1991-01-01

The purpose of this study was to evaluate the effect of probe motion on laser probe temperature in various blood flow conditions. Laser probe temperatures were measured in an in vitro blood circulation model consisting of 3.2 nm-diameter plastic tubes. A 2.0 mm-diameter metal probe attached to a 300 microns optical quartz fiber was coupled to an argon laser. Continuous wave 4 watts and 8 watts of laser power were delivered to the fiber tip corresponding to a 6.7 +/- 0.5 and 13.2 +/- 0.7 watts power setting at the laser generator. The laser probe was either moved with constant velocity or kept stationary. A thermocouple inserted in the lateral portion of the probe was used to record probe temperatures. Probe temperature changes were found with the variation of laser power, probe velocity, blood flow, and duration of laser exposure. Probe motion significantly reduced probe temperatures. After 10 seconds of 4 watts laser power the probe temperature in stagnant blood decreased from 303 +/- 18 degrees C to 113 +/- 17 degrees C (63%) by moving the probe with a velocity of 5 cm/sec. Blood flow rates of 170 ml/min further decreased the probe temperature from 113 +/- 17 degrees C to 50 +/- 8 degrees C (56%). At 8 watts of laser power a probe temperature reduction from 591 +/- 25 degrees C to 534 +/- 36 degrees C (10%) due to 5 cm/sec probe velocity was noted. Probe temperatures were reduced to 130 +/- 30 degrees C (78%) under the combined influence of 5 cm/sec probe velocity and 170 ml/min blood flow.(ABSTRACT TRUNCATED AT 250 WORDS)

Gram stains: a resource for retrospective analysis of bacterial pathogens in clinical studies.

Science.gov (United States)

Srinivasan, Usha; Ponnaluri, Sreelatha; Villareal, Lisa; Gillespie, Brenda; Wen, Ai; Miles, Arianna; Bucholz, Brigette; Marrs, Carl F; Iyer, Ram K; Misra, Dawn; Foxman, Betsy

2012-01-01

We demonstrate the feasibility of using qPCR on DNA extracted from vaginal Gram stain slides to estimate the presence and relative abundance of specific bacterial pathogens. We first tested Gram stained slides spiked with a mix of 10(8) cfu/ml of Escherichia coli and 10(5) cfu/ml of Lactobacillus acidophilus. Primers were designed for amplification of total and species-specific bacterial DNA based on 16S ribosomal gene regions. Sample DNA was pre-amplified with nearly full length 16S rDNA ribosomal gene fragment, followed by quantitative PCR with genera and species-specific 16S rDNA primers. Pre-amplification PCR increased the bacterial amounts; relative proportions of Escherichia coli and Lactobacillus recovered from spiked slides remained unchanged. We applied this method to forty two archived Gram stained slides available from a clinical trial of cerclage in pregnant women at high risk of preterm birth. We found a high correlation between Nugent scores based on bacterial morphology of Lactobacillus, Gardenerella and Mobiluncus and amounts of quantitative PCR estimated genus specific DNA (rrn copies) from Gram stained slides. Testing of a convenience sample of eight paired vaginal swabs and Gram stains freshly collected from healthy women found similar qPCR generated estimates of Lactobacillus proportions from Gram stained slides and vaginal swabs. Archived Gram stained slides collected from large scale epidemiologic and clinical studies represent a valuable, untapped resource for research on the composition of bacterial communities that colonize human mucosal surfaces.

Indocyanine green staining facilitates detection of bleb leakage during trabeculectomy.

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Okazaki, Teruhiko; Kiuchi, Takahiro; Kawana, Keisuke; Oshika, Tetsuro

2007-03-01

To report a new technique to visualize bleb leakage using indocyanine green (ICG) staining during trabeculectomy. The ICG solution was widely applied over the filtering bleb including the conjunctival wound before completion of trabeculectomy. This procedure was performed in 48 eyes of 44 consecutive patients undergoing trabeculectomy between December 2004 and October 2005. Without staining, bleb leakage was not identified by the direct observation under the operating microscope. ICG staining clearly visualized aqueous leakage from the bleb in 5 eyes (10.4%). The bleb leakage in these eyes was easily repaired with 10-0 nylon sutures, and no eyes, including these 5 cases, showed bleb leakage after surgery. There were no intraoperative and postoperative complications related to ICG application. The application of ICG during trabeculectomy is a simple and useful technique to facilitate detection and repair of the bleb leakage.

Mercury localization in mouse kidney over time: autoradiography versus silver staining

International Nuclear Information System (INIS)

Rodier, P.M.; Kates, B.; Simons, R.

1988-01-01

Several methods of silver staining have been employed to localize mercury in tissue, under the assumption that the techniques represent total Hg, but recent reports have suggested that these stains are specific for a limited fraction of the Hg present in some samples. Magos et al. hypothesized that the stains actually vary with inorganic mercury content. The purpose of the present study was to compare localization by radiolabeling to localization by one silver stain, the photoemulsion histochemical technique, in tissues prepared to contain a range of levels of total Hg and a range of levels of inorganic Hg. Mice dosed with 8 mg Hg/kg as MeHg were killed 24 hr, 1 week, or 2 weeks after exposure, to allow a decrease in total Hg and an increase in the proportion of demethylated Hg over time. Mice dosed with 4 mg Hg/kg as HgCl 2 provided samples in which all the Hg present was in the inorganic form. Atomic absorption of kidneys of mice dosed with MeHg showed that total Hg fell from 55 micrograms/g to 39 to 25 over 2 weeks, while the inorganic fraction climbed from about 2 to 27 to 35%. Grain counts from autoradiographs of 203 Hg-labeled sections correlated with total Hg content at +0.88, but silver staining was correlated with inorganic Hg content, appearing only at late termination times in MeHg-exposed animals, but soon after dosing in mice exposed to inorganic Hg. The photoemulsion histochemical technique revealed a substance strictly localized in the proximal tubules, while autoradiographs and grain counts showed total Hg to be present throughout the kidney tissue. These results support the contention that silver stains are selective for inorganic Hg

Embedding, serial sectioning and staining of zebrafish embryos using JB-4 resin.

Science.gov (United States)

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.

Optimal staining methods for delineation of cortical areas and neuron counts in human brains.

Science.gov (United States)

Uylings, H B; Zilles, K; Rajkowska, G

1999-04-01

For cytoarchitectonic delineation of cortical areas in human brain, the Gallyas staining for somata with its sharp contrast between cell bodies and neuropil is preferable to the classical Nissl staining, the more so when an image analysis system is used. This Gallyas staining, however, does not appear to be appropriate for counting neuron numbers in pertinent brain areas, due to the lack of distinct cytological features between small neurons and glial cells. For cell counting Nissl is preferable. In an optimal design for cell counting at least both the Gallyas and the Nissl staining must be applied, the former staining for cytoarchitectural delineaton of cortical areas and the latter for counting the number of neurons in the pertinent cortical areas. Copyright 1999 Academic Press.

The utility of the hematoxylin and eosin staining in patients with suspected Hirschsprung disease.

Science.gov (United States)

Setiadi, Josephine Amanda; Dwihantoro, Andi; Iskandar, Kristy; Heriyanto, Didik Setyo; Gunadi

2017-06-19

While immunohistochemistry (IHC) methods have been widely conducted for the diagnosis of Hirschsprung disease (HSCR) in developed countries, there are very few studies on their use in developing countries where hematoxylin and eosin (HE) staining is a key element of the diagnosis of HSCR. We aimed to determine the accuracy of HE staining in the diagnosis of HSCR using S100 IHC as the reference standard in Indonesia. All histopathology performed for the suspicion of HSCR patients from January 2013 to August 2015 in Dr. Sardjito Hospital, Yogyakarta, Indonesia, were retrospectively reviewed. Our study included 23 HSCR patients: 9 males and 14 females. The HE staining revealed 14 negative (absence of ganglion cells) cases (61%) and 9 positive (presence of ganglion cells) cases (39%). In S100 IHC, out of the 9 positive cases by HE staining, 6 (67%) were confirmed for having ganglion cells; and out of the 14 negative cases by HE staining, 12 (86%) were reported negative and 2 (14%) were positive by S100 IHC staining. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy rates of the HE staining were 80% (95% CI: 0.51-0.95), 75% (95% CI: 0.36-0.96), 85.7% (95% CI: 0.56-0.98), 66.7% (95% CI: 0.31-0.91), and 78.3% (95% CI: 0.58-0.90), respectively. Our study showed that HE staining has relatively moderate accuracy for the diagnosis of HSCR. The use of HE staining is still recommended for the diagnosis of HSCR given the limitation of resource allocation for more expensive IHC technologies in developing countries.

Marine Red Staining of a Pennsylvanian Carbonate Slope: Environmental and Oceanographic Significance

NARCIS (Netherlands)

van der Kooij, B.; Immenhauser, A.M.; Steuber, T; Hagmaier, M.; Bahamonde, J.R.; Samankassou, E.; Merino Tomé, O.

2007-01-01

Red-stained platform facies are a common feature of many carbonate settings throughout the geological record. Although the mechanisms involved in red staining of subaerially exposed or argillaceous, peri-platforin limestones are reasonably well understood, the environmental and oceanographic

Stain-etched porous silicon nanostructures for multicrystalline silicon-based solar cells

Science.gov (United States)

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 μs.

One Small Step for the Gram Stain, One Giant Leap for Clinical Microbiology.

Science.gov (United States)

Thomson, Richard B

2016-06-01

The Gram stain is one of the most commonly performed tests in the clinical microbiology laboratory, yet it is poorly controlled and lacks standardization. It was once the best rapid test in microbiology, but it is no longer trusted by many clinicians. The publication by Samuel et al. (J. Clin. Microbiol. 54:1442-1447, 2016, http://dx.doi.org/10.1128/JCM.03066-15) is a start for those who want to evaluate and improve Gram stain performance. In an age of emerging rapid molecular results, is the Gram stain still relevant? How should clinical microbiologists respond to the call to reduce Gram stain error rates? Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Composition of 12-18th century window glass in Belgium: Non-figurative windows in secular buildings and stained-glass windows in religious buildings

International Nuclear Information System (INIS)

Schalm, Olivier; Janssens, Koen; Wouters, Hilde; Caluwe, Danielle

2007-01-01

A set of ca. 500 window glass fragments originating from different historical sites in Belgium and covering the period 12 th -18 th century was analyzed by means of electron probe microanalysis. Most samples are archaeological finds deriving from non-figurative windows in secular buildings. However, the analyzed set also contains glass sampled from still existing non-figurative windows in secular buildings and stained-glass windows in religious buildings. A sudden compositional change at the end of the 14 th century can be noticed among the series of glass compositions that were obtained. These changes could be related to the use of different glassmaker recipes and to the introduction of new raw materials for glass making

Composition of 12-18 th century window glass in Belgium: Non-figurative windows in secular buildings and stained-glass windows in religious buildings

Science.gov (United States)

Schalm, Olivier; Janssens, Koen; Wouters, Hilde; Caluwé, Danielle

2007-07-01

A set of ca. 500 window glass fragments originating from different historical sites in Belgium and covering the period 12 th-18 th century was analyzed by means of electron probe microanalysis. Most samples are archaeological finds deriving from non-figurative windows in secular buildings. However, the analyzed set also contains glass sampled from still existing non-figurative windows in secular buildings and stained-glass windows in religious buildings. A sudden compositional change at the end of the 14 th century can be noticed among the series of glass compositions that were obtained. These changes could be related to the use of different glassmaker recipes and to the introduction of new raw materials for glass making.

Investigation of black soot staining in houses

Energy Technology Data Exchange (ETDEWEB)

Fugler, D. [Canada Mortgage and Housing Corp., Ottawa, ON (Canada)

2000-07-01

Air quality investigators are frequently called upon to determine the origin of streaking, staining or soot marks in both new and old homes. Those marks display common characteristics: black marks along baseboards at interior or exterior walls, behind furniture and at doorways; black smudges on window frames and plastic cabinets; and even shadowing of studs on exterior wall drywall in a few cases. In most instances, carbon soot from a combustion source is the culprit. The combustion sources include furnaces, water heaters, fireplaces, gas dryers, gas ranges, smoking, vehicle exhaust and candle burning. Scepticism about candle soot is prevalent among callers. As a result, a study was initiated in homes where occupants burn candles regularly to investigate soot problems. Samples were collected from five homes, and included stained carpets, filters, and swab samples of black dust or soot. All the houses selected for the study had been built within a three-year period. Some samples of candles commonly burned in those homes were burnt in a laboratory. Air quality audits had been performed in the homes and had revealed other potential pollutant sources. Best practices for cost-effective clean up and control of soot were researched in industry information. The tests conducted in the laboratory found materials consistent with candle soot or residue during microscopic investigations, but no link was established with the stained material obtained from the homes. A few tips for homeowners were included concerning candle burning, and tips for builders were also offered. 1 tab.

Staining of Platyhelminthes by herbal dyes: An eco-friendly technique for the taxonomist

Directory of Open Access Journals (Sweden)

Niranjan Kumar

2015-11-01

Full Text Available Aim: An environment compatible technique to stain Platyhelminthes, Fasciola gigantica, Gastrothylax crumenifer, Taenia solium, and Moniezia expansa using aqueous and alcoholic extract of sugar beet (Beta vulgaris, China rose (Hibiscus rosasinensis, and red rose (Rosa hybrida were described to minimized the deleterious effects of the synthetic dyes. Materials and Methods: Aqueous/ethanolic extracts of roses were extracted from the flowers while red beet was extracted from the roots. Results: Stained helminthes acquired a comparable level of pigmentation with the distinction of their internal structure in these natural dyes. The flukes (liver and rumen internal structure, oral and ventral/posterior sucker, cirrus sac, gravid uterus, testes, ovary, and vitallaria were appeared pink color in aqueous and alcoholic extract of either China or red rose and yellow to brown color in sugar beet stain. The interior of the proglottid of T. solium and M. expansa took yellow to brown color with good contrast in sugar beet stain and of pink to pink-red in China and red rose stain. Conclusion: The extract of roses (red rose followed by China rose followed by red beet possess the potential to replace the conventional stains in the taxonomic study of Platyhelminthes parasites.

A comparative assessment of commonly employed staining ...

African Journals Online (AJOL)

Following an increase in the number of reports of Cryptosporidium infections and the problems encountered in detecting these organisms in faecal smears, a comparative assessment of a modification of the Sheather's flotation technique and other commonly employed staining procedures proved the modified Sheather's ...

Stain-free histopathology by programmable supercontinuum pulses

DEFF Research Database (Denmark)

Tu, Haohua; Liu, Yuan; Turchinovich, Dmitry

2016-01-01

The preparation, staining, visualization and interpretation of histological images of tissue is well accepted as the gold standard process for the diagnosis of disease. These methods have a long history of development, and are used ubiquitously in pathology, despite being highly time- and labour-...

A novel, modernized Golgi-Cox stain optimized for CLARITY cleared tissue.

Science.gov (United States)

Kassem, Mustafa S; Fok, Sandra Y Y; Smith, Kristie L; Kuligowski, Michael; Balleine, Bernard W

2018-01-15

High resolution neuronal information is extraordinarily useful in understanding the brain's functionality. The development of the Golgi-Cox stain allowed observation of the neuron in its entirety with unrivalled detail. Tissue clearing techniques, e.g., CLARITY and CUBIC, provide the potential to observe entire neuronal circuits intact within tissue and without previous restrictions with regard to section thickness. Here we describe an improved Golgi-Cox stain method, optimised for use with CLARITY and CUBIC that can be used in both fresh and fixed tissue. Using this method, we were able to observe neurons in their entirety within a fraction of the time traditionally taken to clear tissue (48h). We were also able to show for the first-time that Golgi stained tissue is fluorescent when visualized using a multi-photon microscope, allowing us to image synaptic spines with a detail previously unachievable. These novel methods provide cheap and easy to use techniques to investigate the morphology of cellular processes in the brain at a new-found depth, speed, utility and detail, without previous restrictions of time, tissue type and section thickness. This is the first application of a Golgi-Cox stain to cleared brain tissue, it is investigated and discussed in detail, describing different methodologies that may be used, a comparison between the different clearing techniques and lastly the novel interaction of these techniques with this ultra-rapid stain. Copyright © 2017 Elsevier B.V. All rights reserved.

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

Science.gov (United States)

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.

Identification of blue staining vaccine-derived material in inflammatory lesions using cultured canine macrophages.

Science.gov (United States)

Scruggs, Jennifer L; LeBlanc, Casey J

2015-03-01

Vaccine reactions are described in cytology textbooks as having eosinophilic to magenta colored globules within and admixed with inflammatory cells. Recently, we have seen increased numbers of inflammatory lesions containing blue to blue-gray globular material, with historical information suggesting an association with rabies vaccination. The purpose of the study was to confirm the blue-gray and the eosinophilic material observed microscopically in some inflammatory lesions as being vaccine-derived. Three different vaccines were cytocentrifuged and Wright stained. Vaccine aliquots were also added to the culture media of canine-derived macrophages for 24 hours and the cells subsequently harvested, cytocentrifuged, and Wright stained. The globular material present in both preparations was compared to that observed in vaccine-induced inflammatory lesions. Morin staining was used to identify metal within vaccine material in both in vitro- and in vivo-derived cytology samples. Vaccine-derived material has a characteristic color and appearance. Appearance of the material was consistent in cytologic samples, in cells incubated with the vaccine, and in cytocentrifuged preparations of the vaccine vial contents. The blue-gray globules stained positively for Morin stain, while the eosinophilic material did not stain. Vaccine-induced inflammatory lesions may contain blue to blue-gray or magenta stained globular material. Blue-gray material was associated with administration of rabies vaccine Imrab 3 TF and the observed material may be metal-containing adjuvant. Magenta material was associated with other vaccines and negative for Morin stain, suggesting a metal-free adjuvant. © 2014 American Society for Veterinary Clinical Pathology.

Development of a preparation and staining method for fetal erythroblasts in maternal blood : Simultaneous immunocytochemical staining and FISH analysis

NARCIS (Netherlands)

Oosterwijk, JC; Mesker, WE; Ouwerkerk-van Velzen, MCM; Knepfle, CFHM; Wiesmeijer, KC; van den Burg, MJM; Beverstock, GC; Bernini, LF; van Ommen, Gert-Jan B; Kanhai, HHH; Tanke, HJ

1998-01-01

In order to detect fetal nucleated red blood cells (NRBCs) in maternal blood, a protocol was developed which aimed at producing a reliable staining method for combined immunocytochemical and FISH analysis. The technique had to be suitable for eventual automated screening of slides. Chorionic villi

The Role of Hemiwicking on the Shape of a Blood Drop Stain

Science.gov (United States)

Shiri, Samira; Martin, Kenneth; Bird, James

2017-11-01

Blood pattern analysis (BPA) typically assumes that an elliptical stain is due to oblique drop impact. From the eccentricity of the elliptical stain - while also accounting for gravity and drag - the source and trajectory of the blood drops can be estimated. Yet, these models generally neglect any fluid motion following impact that could influence the shape of the stain. Here we demonstrate that under certain conditions on certain materials, a blood drop will undergo anisotropic hemiwicking. Through systemic experiments and modeling, we aim to better understand this phenomenon with the goal of ultimately decreasing the uncertainty in crime scene reconstruction.

Temporal variation of VOC emission from solvent and water based wood stains

Science.gov (United States)

de Gennaro, Gianluigi; Loiotile, Annamaria Demarinis; Fracchiolla, Roberta; Palmisani, Jolanda; Saracino, Maria Rosaria; Tutino, Maria

2015-08-01

Solvent- and water-based wood stains were monitored using a small test emission chamber in order to characterize their emission profiles in terms of Total and individual VOCs. The study of concentration-time profiles of individual VOCs enabled to identify the compounds emitted at higher concentration for each type of stain, to examine their decay curve and finally to estimate the concentration in a reference room. The solvent-based wood stain was characterized by the highest Total VOCs emission level (5.7 mg/m3) that decreased over time more slowly than those related to water-based ones. The same finding was observed for the main detected compounds: Benzene, Toluene, Ethylbenzene, Xylenes, Styrene, alpha-Pinene and Camphene. On the other hand, the highest level of Limonene was emitted by a water-based wood stain. However, the concentration-time profile showed that water-based product was characterized by a remarkable reduction of the time of maximum and minimum emission: Limonene concentration reached the minimum concentration in about half the time compared to the solvent-based product. According to AgBB evaluation scheme, only one of the investigated water-based wood stains can be classified as a low-emitting product whose use may not determine any potential adverse effect on human health.

Juvenile localized scleroderma with port wine stain: Coincidental or possible common pathogenetic association

Directory of Open Access Journals (Sweden)

Seval Dogruk Kacar

2015-01-01

Full Text Available 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.

The usefulness of changing focus during examination using Gram staining as initial diagnostic clue for infective tuberculosis.

Science.gov (United States)

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.

An optimized staining technique for the detection of Gram positive and Gram negative bacteria within tissue.

Science.gov (United States)

Becerra, Sandra C; Roy, Daniel C; Sanchez, Carlos J; Christy, Robert J; Burmeister, David M

2016-04-12

Bacterial infections are a common clinical problem in both acute and chronic wounds. With growing concerns over antibiotic resistance, treatment of bacterial infections should only occur after positive diagnosis. Currently, diagnosis is delayed due to lengthy culturing methods which may also fail to identify the presence of bacteria. While newer costly bacterial identification methods are being explored, a simple and inexpensive diagnostic tool would aid in immediate and accurate treatments for bacterial infections. Histologically, hematoxylin and eosin (H&E) and Gram stains have been employed, but are far from optimal when analyzing tissue samples due to non-specific staining. The goal of the current study was to develop a modification of the Gram stain that enhances the contrast between bacteria and host tissue. A modified Gram stain was developed and tested as an alternative to Gram stain that improves the contrast between Gram positive bacteria, Gram negative bacteria and host tissue. Initially, clinically relevant strains of Pseudomonas aeruginosa and Staphylococcus aureus were visualized in vitro and in biopsies of infected, porcine burns using routine Gram stain, and immunohistochemistry techniques involving bacterial strain-specific fluorescent antibodies as validation tools. H&E and Gram stain of serial biopsy sections were then compared to a modification of the Gram stain incorporating a counterstain that highlights collagen found in tissue. The modified Gram stain clearly identified both Gram positive and Gram negative bacteria, and when compared to H&E or Gram stain alone provided excellent contrast between bacteria and non-viable burn eschar. Moreover, when applied to surgical biopsies from patients that underwent burn debridement this technique was able to clearly detect bacterial morphology within host tissue. We describe a modification of the Gram stain that provides improved contrast of Gram positive and Gram negative microorganisms within host

Weathering effects on materials from historical stained glass windows

Directory of Open Access Journals (Sweden)

García-Heras, M.

2003-06-01

Full Text Available A selection of materials (stained glasses, lead cames, support elements and putty from historical stained glass windows of different periods (13th-19th centuries have been studied. Optical microscopy, scanning electron microscopy, energy dispersive X-ray spectrometry and X-ray diffraction were used as characterization techniques. Degradation of historical stained glass windows is due to the particular chemical composition oftlie materials used for their production: stained glasses, lead network, metallic support elements and refilling putty. However, the presence of a given chemical composition is not the only factor involved in the degradation process. It is necessary the occurrence of other external factors that contribute to the development and progress of alteration problems in the materials mentioned above. The presence of gaseous pollution in the air produces a negative interaction with the surface of the stained glass windows materials. Firstly, the stained glasses and the grisailles begin a dealkalinisation process and a silica gel layer is formed during the early contact between the glasses and the wet environment. After that, insoluble salt deposits and corrosion crusts are formed as a consequence of a deeper chemical attack which results in a depolymerisation of the glass network. The lead cames and the metallic support elements are also altered by weathering. Such materials are oxidized and both pits and crusts appear on their surfaces. The transport of ions and other substances from the corrosion crusts of the metallic elements gives rise new deposits upon the stained glasses, which could intensify their own degradation processes. The putty experiments a noticeable shrinkage and cracking. Likewise, adverse environmental conditions favour the transport of putty substances towards the other materials of the stained glass window, thereby increasing the crusts thickness and adding elements that contribute to the total alteration of the

Teeth re-whitening effect of strawberry juice on coffee stained teeth

Directory of Open Access Journals (Sweden)

Annisya Pramesti

2018-01-01

Full Text Available Many people favor coffee. However, regarding health and aesthetic dentistry, coffee gives a negative effect. Tanin in coffee causes a brown stain on the tooth surface. Therefore, in aesthetic dental care, teeth whitening has become popular matter. One of the natural ingredients used for teeth whitening treatment is strawberry. The purpose of this study was to obtained data regarding the effect of strawberry juice on the re-whitening process of the coffee-stained tooth enamel surface. This study was a pure experimental in-vitro using Friedman and Wilcoxon Matched Pairs Tests for statistical analysis. The population of this study was anterior teeth. The samples were maxillary central incisors. The sampling technique using sample size determination based on the testing formulas of the difference of two average data pairs resulted in 11 specimens. The result of the research showed that all coffee-stained teeth sample had an increasing enamel colour index. The samples were then applied with strawberry juice resulted in a significant average difference colour index value indicated by p<0.001. The conclusion of this research indicated that there was an effect of strawberry juice on the coffee-stained teeth re-whitening process.

WITHDRAWN: Amnioinfusion for meconium-stained liquor in labour.

Science.gov (United States)

Hofmeyr, G Justus

2009-01-21

Amnioinfusion aims to prevent or relieve umbilical cord compression during labour by infusing a solution into the uterine cavity. It is also thought to dilute meconium when present in the amniotic fluid and so reduce the risk of meconium aspiration. However, it may be that the mechanism of effect is that it corrects oligohydramnios (reduced amniotic fluid), for which thick meconium staining is a marker. The objective of this review was to assess the effects of amnioinfusion for meconium-stained liquor on perinatal outcome. The Cochrane Pregnancy and Childbirth Group trials register (October 2001) and the Cochrane Controlled Trials Register (Issue 3, 2001) were searched. Randomised trials comparing amnioinfusion with no amnioinfusion for women in labour with moderate or thick meconium-staining of the amniotic fluid. Eligibility and trial quality were assessed by one reviewer. Twelve studies, most involving small numbers of participants, were included. Under standard perinatal surveillance, amnioinfusion was associated with a reduction in the following: heavy meconium staining of the liquor (relative risk 0.03, 95% confidence interval 0.01 to 0.15); variable fetal heart rate deceleration (relative risk 0.65, 95% confidence interval 0.49 to 0.88); and reduced caesarean section overall (relative risk 0.82, 95% confidence interval 0.69 to 1.97). No perinatal deaths were reported. Under limited perinatal surveillance, amnioinfusion was associated with a reduction in the following: meconium aspiration syndrome (relative risk 0.24, 95% confidence interval 0.12 to 0.48); neonatal hypoxic ischaemic encephalopathy (relative risk 0.07, 95% confidence interval 0.01 to 0.56) and neonatal ventilation or intensive care unit admission (relative risk 0.56, 95% confidence interval 0.39 to 0.79); there was a trend towards reduced perinatal mortality (relative risk 0.34, 95% confidence interval 0.11 to 1.06). Amnioinfusion is associated with improvements in perinatal outcome

Prophylactic cefazolin in amnioinfusions administered for meconium-stained amniotic fluid.

Science.gov (United States)

Edwards, R K; Duff, P

1999-01-01

OBJECTIVE: To determine if amnioinfusion with an antibiotic solution decreased the rate of clinical chorioamnionitis and puerperal endometritis in patients with meconium-stained amniotic fluid. METHODS: Patients in labor at 36 weeks of gestation or greater with singleton pregnancies and meconium-stained amniotic fluid were randomized to receive either cefazolin, 1 g/1,000 mL, of normal saline (n = 90) or normal saline (n = 93) amnioinfusion. Rates of clinically diagnosed chorioamnionitis and endometritis and of suspected and culture-proven neonatal infection were determined. RESULTS: Between the study and control groups, the incidences of clinical chorioamnionitis (7.8% vs. 8.6%), endometritis (2.4% vs. 3.5%), aggregate intrauterine infection (10.0% vs. 11.8%), suspected neonatal infection (17.8% vs. 21.5%), and proven neonatal infection (0.0% vs. 2.2%) were not significantly different. CONCLUSIONS: Prophylactic use of cefazolin in amnioinfusions did not significantly reduce rates of maternal or neonatal infection in patients with meconium-stained amniotic fluid. PMID:10371474

Morphological characteristics of developmental stages of Acanthamoeba and Naegleria species before and after staining by various techniques.

Science.gov (United States)

Ithoi, Init; Ahmad, Arine-Fadzlun; Mak, J W; Nissapatorn, Veeranoot; Lau, Yee-Ling; Mahmud, Rohela

2011-11-01

Seven stains were studied to determine the best color and contrast for staining the developmental stages of free living pathogenic Acanthamoeba and Naegleria species. The acid-fast bacilli stain (AFB) produced a blue color without contrast; trichrome-eosin and modified Field's showed various color contrasts; Giemsa, iron-hematoxylin, modified AFB and Gram produced only one color which distinguished the nucleus, nucleolus, cytoplasm, food- and water-vacuoles. The motile organs (acanthopodia, pseudopodia, lobopodia and flagella) were also clearly differentiated but produced a similar color as the cytoplasm. These motile organelles were first induced by incubating at 37 degrees C for at least 15 minutes and then fixing with methanol in order to preserve the protruding morphology prior to staining. The trichrome-eosin and iron-hematoxylin stains showed good color contrast for detecting all three stages, the trophozoite, cyst and flagellate; Giemsa and Gram stained the trophozoite and flagellate stages; the modified Field's and modified AFB stains stained only the trophozoite stage. Depending on the purpose, all these stains (except the AFB stain) can be used to identify the developmental stages of Acanthamoeba and Naegleria for clinical, epidemiological or public health use.

Microscopic analysis of MTT stained boar sperm cells

African Journals Online (AJOL)

tulyasys

2015-06-08

2H-tetrazolium bromide is widely used for assessment of cytotoxicity, cell viability, and proliferation studies in cell biology (van Meerloo et al., 2011;. Stockert et al., 2012). The stain is abbreviated as MTT.

Propidium iodide (PI) stains Nissl bodies and may serve as a quick marker for total neuronal cell count.

Science.gov (United States)

Niu, Junfei; Li, Chunman; Wu, Haihui; Feng, Xianling; Su, Qingning; Li, Shihe; Zhang, Lihong; Yew, David Tai Wai; Cho, Eric Yu Pang; Sha, Ou

2015-03-01

Propidium iodide (PI) reacts with both DNA and RNA and is a commonly used fluorescent reagent for nucleic acid staining. The aim of the study was to compare the cellular staining patterns of PI with that of Nissl staining in rat nervous tissues and to report a modified staining method that selectively labels Nissl bodies in neurons. Cryosections and paraffin sections of different tissues of normal Sprague-Dawley rats, including trigeminal ganglia, dorsal root ganglia, spinal cord, liver, and small intestine, were stained by either PI or the hematoxylin and eosin method. Some sections were treated with RNase or DNase before the above staining, and some were double stained with PI and a Nissl stain. The sections were observed by light, fluorescence or confocal microscopy. Results showed strong PI signals detected as patterns of granules in the neuronal cytoplasm of all nervous tissues, whereas the staining of neuronal nuclei was weaker. In contrast, nuclei of neuroglial cells were strongly stained by PI, while the cytoplasm was not obviously stained. Pretreatment of the neural tissue with RNase abolished the PI signals. Furthermore, the PI positive granules in neuronal cytoplasm co-localized with Nissl bodies stained by the fluorescent Nissl stain. When the tissue was pretreated with DNase, PI only stained the cytoplasmic granules of neurons, but not that of glial cells. Our results show that PI stains Nissl bodies and may serve as an economical and convenient neuron marker for neuronal cell counting when specific neural markers such as antibodies are not readily available. Copyright © 2015. Published by Elsevier GmbH.

Four-probe measurements with a three-probe scanning tunneling microscope

International Nuclear Information System (INIS)

Salomons, Mark; Martins, Bruno V. C.; Zikovsky, Janik; Wolkow, Robert A.

2014-01-01

We present an ultrahigh vacuum (UHV) three-probe scanning tunneling microscope in which each probe is capable of atomic resolution. A UHV JEOL scanning electron microscope aids in the placement of the probes on the sample. The machine also has a field ion microscope to clean, atomically image, and shape the probe tips. The machine uses bare conductive samples and tips with a homebuilt set of pliers for heating and loading. Automated feedback controlled tip-surface contacts allow for electrical stability and reproducibility while also greatly reducing tip and surface damage due to contact formation. The ability to register inter-tip position by imaging of a single surface feature by multiple tips is demonstrated. Four-probe material characterization is achieved by deploying two tips as fixed current probes and the third tip as a movable voltage probe

Four-probe measurements with a three-probe scanning tunneling microscope

Energy Technology Data Exchange (ETDEWEB)

Salomons, Mark [National Institute for Nanotechnology, National Research Council of Canada, Edmonton, Alberta T6G 2M9 (Canada); Martins, Bruno V. C.; Zikovsky, Janik; Wolkow, Robert A., E-mail: rwolkow@ualberta.ca [National Institute for Nanotechnology, National Research Council of Canada, Edmonton, Alberta T6G 2M9 (Canada); Department of Physics, University of Alberta, Edmonton, Alberta T6G 2E1 (Canada)

2014-04-15

We present an ultrahigh vacuum (UHV) three-probe scanning tunneling microscope in which each probe is capable of atomic resolution. A UHV JEOL scanning electron microscope aids in the placement of the probes on the sample. The machine also has a field ion microscope to clean, atomically image, and shape the probe tips. The machine uses bare conductive samples and tips with a homebuilt set of pliers for heating and loading. Automated feedback controlled tip-surface contacts allow for electrical stability and reproducibility while also greatly reducing tip and surface damage due to contact formation. The ability to register inter-tip position by imaging of a single surface feature by multiple tips is demonstrated. Four-probe material characterization is achieved by deploying two tips as fixed current probes and the third tip as a movable voltage probe.

Four-probe measurements with a three-probe scanning tunneling microscope.

Science.gov (United States)

Salomons, Mark; Martins, Bruno V C; Zikovsky, Janik; Wolkow, Robert A

2014-04-01

We present an ultrahigh vacuum (UHV) three-probe scanning tunneling microscope in which each probe is capable of atomic resolution. A UHV JEOL scanning electron microscope aids in the placement of the probes on the sample. The machine also has a field ion microscope to clean, atomically image, and shape the probe tips. The machine uses bare conductive samples and tips with a homebuilt set of pliers for heating and loading. Automated feedback controlled tip-surface contacts allow for electrical stability and reproducibility while also greatly reducing tip and surface damage due to contact formation. The ability to register inter-tip position by imaging of a single surface feature by multiple tips is demonstrated. Four-probe material characterization is achieved by deploying two tips as fixed current probes and the third tip as a movable voltage probe.

The effect of corrosion on stained glass windows

Directory of Open Access Journals (Sweden)

Laissner, Johanna

1996-06-01

Full Text Available Stained glass windows belong to the most important cultural heritage of Europe. Within the last decades a disastrous deterioration took place. The wonderful stained glass windows and their glass paintings as pieces of art are acutely menaced by environmental corrosive influences. This corrosion process is a very complex reaction which is not only influenced by temperature and humidity changes but also by gaseous pollutants like sulfur dioxide, nitrogen oxides or ozone, by dust and air, microorganisms as well as synergetic interactions. Strongly affected by these environmental attacks are medieval stained glasses due to their chemical composition. They have a low content in silica and high contents of modifier ions (e.g. potassium and calcium. The corrosion phenomena can range from predominantly pitting on the surface to the formation of thick corrosion crusts which are turning the panel opaque and thus reducing strongly the transparency of the windows. In order to set up a conservation and restoration concept, it is necessary to know about the environmental conditions to which the stained glass windows are exposed. For this purpose very corrosion sensitive model glasses (so called glass sensors were developed which have a similar chemical composition as historic stained glasses. They exhibit the same corrosion reactions but react much faster, and are now widely used to estimate corrosive stresses on stained glass windows to give basic information about the corrosive impacts which work on the historic glasses. In this paper principle corrosion mechanisms of stained glass windows and their enhancing factors are discussed. For the evaluation of the environmental impact, the application of glass sensors is demonstrated.

Las vidrieras coloreadas pertenecen al legado cultural más importante de Europa. En las últimas décadas se ha producido en ellas un desastroso deterioro. Las maravillosas vidrieras coloreadas y sus policromías est

Proposals for best-quality immunohistochemical staining of paraffin-embedded brain tissue slides in forensics.

Science.gov (United States)

Trautz, Florian; Dreßler, Jan; Stassart, Ruth; Müller, Wolf; Ondruschka, Benjamin

2018-01-03

Immunohistochemistry (IHC) has become an integral part in forensic histopathology over the last decades. However, the underlying methods for IHC vary greatly depending on the institution, creating a lack of comparability. The aim of this study was to assess the optimal approach for different technical aspects of IHC, in order to improve and standardize this procedure. Therefore, qualitative results from manual and automatic IHC staining of brain samples were compared, as well as potential differences in suitability of common IHC glass slides. Further, possibilities of image digitalization and connected issues were investigated. In our study, automatic staining showed more consistent staining results, compared to manual staining procedures. Digitalization and digital post-processing facilitated direct analysis and analysis for reproducibility considerably. No differences were found for different commercially available microscopic glass slides regarding suitability of IHC brain researches, but a certain rate of tissue loss should be expected during the staining process.

Probe-based confocal laser endomicroscopy (pCLE) - a new imaging technique for in situ localization of spermatozoa.

Science.gov (United States)

Trottmann, Matthias; Stepp, Herbert; Sroka, Ronald; Heide, Michael; Liedl, Bernhard; Reese, Sven; Becker, Armin J; Stief, Christian G; Kölle, Sabine

2015-05-01

In azoospermic patients, spermatozoa are routinely obtained by testicular sperm extraction (TESE). However, success rates of this technique are moderate, because the site of excision of testicular tissue is determined arbitrarily. Therefore the aim of this study was to establish probe-based laser endomicroscopy (pCLE) a noval biomedical imaging technique, which provides the opportunity of non-invasive, real-time visualisation of tissue at histological resolution. Using pCLE we clearly visualized longitudinal and horizontal views of the tubuli seminiferi contorti and localized vital spermatozoa. Obtained images and real-time videos were subsequently compared with confocal laser scanning microscopy (CLSM) of spermatozoa and tissues, respectively. Comparative visualization of single native Confocal laser scanning microscopy (CLSM, left) and probe-based laser endomicroscopy (pCLE, right) using Pro Flex(TM) UltraMini O after staining with acriflavine. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Staining Methods for Normal and Regenerative Myelin in the Nervous System.

Science.gov (United States)

Carriel, Víctor; Campos, Antonio; Alaminos, Miguel; Raimondo, Stefania; Geuna, Stefano

2017-01-01

Histochemical techniques enable the specific identification of myelin by light microscopy. Here we describe three histochemical methods for the staining of myelin suitable for formalin-fixed and paraffin-embedded materials. The first method is conventional luxol fast blue (LFB) method which stains myelin in blue and Nissl bodies and mast cells in purple. The second method is a LBF-based method called MCOLL, which specifically stains the myelin as well the collagen fibers and cells, giving an integrated overview of the histology and myelin content of the tissue. Finally, we describe the osmium tetroxide method, which consist in the osmication of previously fixed tissues. Osmication is performed prior the embedding of tissues in paraffin giving a permanent positive reaction for myelin as well as other lipids present in the tissue.

[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].

Science.gov (United States)

Szeremeta, Michał; Pepiński, Witold; Niemcunowicz-Janica, Anna; Skawrońska, Małgorzata; Sackiewicz, Adam; Ptaszyńska-Sarosiek, Iwona; Okłota, 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.

Gram staining of protected pulmonary specimens in the early diagnosis of ventilator-associated pneumonia.

Science.gov (United States)

Mimoz, O; Karim, A; Mazoit, J X; Edouard, A; Leprince, S; Nordmann, P

2000-11-01

We evaluated prospectively the use of Gram staining of protected pulmonary specimens to allow the early diagnosis of ventilator-associated pneumonia (VAP), compared with the use of 60 bronchoscopic protected specimen brushes (PSB) and 126 blinded plugged telescopic catheters (PTC) obtained from 134 patients. Gram stains were from Cytospin slides; they were studied for the presence of microorganisms in 10 and 50 fields by two independent observers and classified according to their Gram stain morphology. Quantitative cultures were performed after serial dilution and plating on appropriate culture medium. A final diagnosis of VAP, based on a culture of > or = 10(3) c.f.u. ml-1, was established after 81 (44%) samplings. When 10 fields were analysed, a strong relationship was found between the presence of bacteria on Gram staining and the final diagnosis of VAP (for PSB and PTC respectively: sensitivity 74 and 81%, specificity 94 and 100%, positive predictive value 91 and 100%, negative predictive value 82 and 88%). The correlation was less when we compared the morphology of microorganisms observed on Gram staining with those of bacteria obtained from quantitative cultures (for PSB and PTC respectively: sensitivity 54 and 69%, specificity 86 and 89%, positive predictive value 72 and 78%, negative predictive value 74 and 84%). Increasing the number of fields read to 50 was associated with a slight decrease in specificity and positive predictive value of Gram staining, but with a small increase in its sensitivity and negative predictive value. The results obtained by the two observers were similar to each other for both numbers of fields analysed. Gram staining of protected pulmonary specimens performed on 10 fields predicted the presence of VAP and partially identified (using Gram stain morphology) the microorganisms growing at significant concentrations, and could help in the early choice of the treatment of VAP. Increasing the number of fields read or having the Gram

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

International Nuclear Information System (INIS)

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

Gram and acridine orange staining for diagnosis of septic arthritis in different patient populations.

Science.gov (United States)

Cunningham, Gregory; Seghrouchni, Khalid; Ruffieux, Etienne; Vaudaux, Pierre; Gayet-Ageron, Angèle; Cherkaoui, Abdessalam; Godinho, Eduardo; Lew, Daniel; Hoffmeyer, Pierre; Uçkay, Ilker

2014-06-01

The sensitivity of Gram staining is known to be suboptimal for the diagnosis of native joint septic arthritis. We lack information about the accuracy of Gram compared to other microscopic staining techniques for predicting infection in different patient populations. This was a cohort study with cost evaluations at the Orthopaedic Service of Geneva University Hospitals (January 1996-October 2012). Among 500 episodes of arthritis (196 with immunosuppression, 227 with underlying arthroplasties and 69 with gout or other crystals in synovial fluid), Gram staining revealed pathogens in 146 episodes (146/500, 29 %) or in 146 of the 400 culture-positive episodes (37 %). Correlation between the Gram and acridine staining of the same sample was good (Spearman 0.85). Overall, the sensitivity, specificity, positive predictive value and negative predictive value of Gram stain for rapid diagnosis of septic arthritis was 0.37, 0.99, 0.99 and 0.28, respectively, compared to microbiological cultures. Quite similar values were recorded across the different patient subpopulations, in particular for sensitivity values that were 0.33 for patients with prosthetic joint infections, 0.40 for immunosuppressed patients, 0.36 for patients under antibiotic administration and 0.52 for patients with concomitant crystalline disease. The sensitivity of Gram or acridine orange staining for a rapid diagnosis of episodes of septic arthritis is suboptimal compared to microbiological culture, regardless of underlying conditions, immunosuppression or antibiotic therapy. The sensitivity in the presence of synovial fluid crystals is moderate. Acridine orange and Gram stains are equivalent.

A mitochondrial targeted two-photon iridium(III) phosphorescent probe for selective detection of hypochlorite in live cells and in vivo.

Science.gov (United States)

Li, Guanying; Lin, Qian; Sun, Lingli; Feng, Changsheng; Zhang, Pingyu; Yu, Bole; Chen, Yu; Wen, Ya; Wang, Hui; Ji, Liangnian; Chao, Hui

2015-01-01

Endogenous hypochlorite ion (ClO(-)) is a highly reactive oxygen species (ROS) that is produced from hydrogen peroxide and chloride ions catalyzed by myeloperoxidase (MPO). And mitochondrion is one of the major sources of ROS including ClO(-). In the present work, a two-photon phosphorescent probe for ClO(-) in mitochondria was developed. An iridium(III) complex bearing a diaminomaleonitrile group as ClO(-) reactive moiety specifically responded to ClO(-) over other ions and ROSs. When the probe was reacted with ClO(-) to form an oxidized carboxylate product, a significant enhancement in phosphorescence intensity was observed under one-photon (402 nm) and two-photon (750 nm) excitation, with a two-photon absorption cross-section of 78.1 GM at 750 nm. More importantly, ICP-MS results and cellular images co-stained with Mito-tracker Green demonstrated that this probe possessed high specificity for mitochondria. This probe was applied in the one- and two-photon imaging of ClO(-) in vitro and in vivo. The results suggested endotoxin lipopolysaccharide (LPS) induced ClO(-) mostly generated in the liver of zebrafish. Copyright © 2015 Elsevier Ltd. All rights reserved.

Mobile Game Probes

DEFF Research Database (Denmark)

Borup Lynggaard, Aviaja

2006-01-01

This paper will examine how probes can be useful for game designers in the preliminary phases of a design process. The work is based upon a case study concerning pervasive mobile phone games where Mobile Game Probes have emerged from the project. The new probes are aimed towards a specific target...... group and the goal is to specify the probes so they will cover the most relevant areas for our project. The Mobile Game Probes generated many interesting results and new issues occurred, since the probes came to be dynamic and favorable for the process in new ways....

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

Science.gov (United States)

Ginsberg, Stephen D; Che, Shaoli

2004-08-01

The use of five histochemical stains (cresyl violet, thionin, hematoxylin & eosin, silver stain, and acridine orange) was evaluated in combination with an expression profiling paradigm that included regional and single cell analyses within the hippocampus of post-mortem human brains and adult mice. Adjacent serial sections of human and mouse hippocampus were labeled by histochemistry or neurofilament immunocytochemistry. These tissue sections were used as starting material for regional and single cell microdissection followed by a newly developed RNA amplification procedure (terminal continuation (TC) RNA amplification) and subsequent hybridization to custom-designed cDNA arrays. Results indicated equivalent levels of global hybridization signal intensity and relative expression levels for individual genes for hippocampi stained by cresyl violet, thionin, and hematoxylin & eosin, and neurofilament immunocytochemistry. Moreover, no significant differences existed between the Nissl stains and neurofilament immunocytochemistry for individual CA1 neurons obtained via laser capture microdissection. In contrast, a marked decrement was observed in adjacent hippocampal sections stained for silver stain and acridine orange, both at the level of the regional dissection and at the CA1 neuron population level. Observations made on the cDNA array platform were validated by real-time qPCR using primers directed against beta-actin and glyceraldehyde-3 phosphate dehydrogenase. Thus, this report demonstrated the utility of using specific Nissl stains, but not stains that bind RNA species directly, in both human and mouse brain tissues at the regional and cellular level for state-of-the-art molecular fingerprinting studies.

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

Energy Technology Data Exchange (ETDEWEB)

Delgado, J. [Dep. de Conservacao e Restauro, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); Vilarigues, M. [Dep. de Conservacao e Restauro, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); VICARTE, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); Ruivo, A. [VICARTE, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); REQUIMTE, FCT-UNL, Quinta da Torre, 2829-516 Caparica (Portugal); Corregidor, V.; Silva, R.C. da [Unidade de Fisica e Aceleradores, LFI, ITN, E.N.10, 2686-953 Sacavem (Portugal); CFNUL, Av., Prof. Gama Pinto n 2, 1649-003 Lisboa (Portugal); Alves, L.C., E-mail: lcalves@itn.pt [Unidade de Fisica e Aceleradores, LFI, ITN, E.N.10, 2686-953 Sacavem (Portugal); CFNUL, Av., Prof. Gama Pinto n 2, 1649-003 Lisboa (Portugal)

2011-10-15

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 {sup o}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.

Differences in staining intensities affect reported occurrences and concentrations of Giardia spp. in surface drinking water sources.

Science.gov (United States)

Alderisio, K A; Villegas, L F; Ware, M W; McDonald, L A; Xiao, L; Villegas, E N

2017-12-01

USEPA Method 1623, or its equivalent, is currently used to monitor for protozoan contamination of surface drinking water sources worldwide. At least three approved staining kits used for detecting Cryptosporidium and Giardia are commercially available. This study focuses on understanding the differences among staining kits used for Method 1623. Merifluor and EasyStain labelling kits were used to monitor Cryptosporidium oocyst and Giardia cyst densities in New York City's raw surface water sources. In the year following a change to the approved staining kits for use with Method 1623, an anomaly was noted in the occurrence of Giardia cysts in New York City's raw surface water. Specifically, Merifluor-stained samples had higher Giardia cyst densities as compared with those stained with EasyStain. Side by side comparison revealed significantly lower fluorescence intensities of Giardia muris as compared with Giardia duodenalis cysts when labelled with EasyStain. This study showed very poor fluorescence intensity signals by EasyStain on G. muris cysts resulting in lower cyst counts, while Merifluor, with its broader Giardia cyst staining specificity, resulted in higher cyst counts, when using Methods 1623. These results suggest that detected Giardia cyst concentrations are dependent on the staining kits used, which can result in a more or less conservative estimation of occurrences and densities of zoonotic Giardia cysts by detecting a broader range of Giardia species/Assemblages. Published 2017. This article is a U.S. Government work and is in the public domain in the USA.

Gloss and Stain Resistance of Ceramic-Polymer CAD/CAM Restorative Blocks.

Science.gov (United States)

Lawson, Nathaniel C; Burgess, John O

2016-03-01

To evaluate the gloss and stain resistance of several new ceramic-polymer CAD/CAM blocks Specimens (4 mm) were sectioned from: Enamic (polymer-infused ceramic), LAVA Ultimate (nano-ceramic reinforced polymer), e.max (lithium disilicate), Paradigm C (porcelain), and Paradigm MZ100 (composite). Specimens were wet polished on a polishing wheel to either 320 grit silicon paper (un-polished, N = 8) or 2000 grit silicon carbide papers followed by a 0.05 μm alumina slurry (polished, N = 8). Initial gloss and color (L*a*b*) values were measured. Specimens were stored in a staining solution at 37°C in darkness for 12 days (simulating 1 year). After storage, L*a*b* values re-measured. Change in color was reported as ΔE00 based on the CIEDE2000 formula. Gloss and ΔE00 were analyzed by two-way analysis of variance (ANOVA) (alpha = .05). Separate one-way ANOVA and Tukey post-hoc analyses were performed for both polish conditions and all materials. Two-way ANOVA showed that factors material, polish and their interaction were significant for both gloss and ΔE00 (p gloss and less color change than all other materials. The composition and polish of CAD/CAM materials affects gloss and stain resistance. Ceramic-polymer hybrid materials can achieve the high gloss required for esthetic restorations. These materials should be polished in order to minimize staining. If polished, all of the tested materials exhibited clinically acceptable color changes at 1 year of simulated staining. (J Esthet Restor Dent 28:S40-S45, 2016). © 2015 Wiley Periodicals, Inc.

Traversing probe system

Science.gov (United States)

Mashburn, Douglas N.; Stevens, Richard H.; Woodall, Harold C.

1977-01-01

This invention comprises a rotatable annular probe-positioner which carries at least one radially disposed sensing probe, such as a Pitot tube having a right-angled tip. The positioner can be coaxially and rotatably mounted within a compressor casing or the like and then actuated to orient the sensing probe as required to make measurements at selected stations in the annulus between the positioner and compressor casing. The positioner can be actuated to (a) selectively move the probe along its own axis, (b) adjust the yaw angle of the right-angled probe tip, and (c) revolve the probe about the axis common to the positioner and casing. A cam plate engages a cam-follower portion of the probe and normally rotates with the positioner. The positioner includes a first-motor-driven ring gear which effects slidable movement of the probe by rotating the positioner at a time when an external pneumatic cylinder is actuated to engage the cam plate and hold it stationary. When the pneumatic cylinder is not actuated, this ring gear can be driven to revolve the positioner and thus the probe to a desired circumferential location about the above-mentioned common axis. A second motor-driven ring gear included in the positioner can be driven to rotate the probe about its axis, thus adjusting the yaw angle of the probe tip. The positioner can be used in highly corrosive atmosphere, such as gaseous uranium hexafluoride.

Traversing probe system

International Nuclear Information System (INIS)

Mashburn, D.N.; Stevens, R.H.; Woodall, H.C.

1977-01-01

This invention comprises a rotatable annular probe-positioner which carries at least one radially disposed sensing probe, such as a Pitot tube having a right-angled tip. The positioner can be coaxially and rotatably mounted within a compressor casing or the like and then actuated to orient the sensing probe as required to make measurements at selected stations in the annulus between the positioner and compressor casing. The positioner can be actuated to (a) selectively move the probe along its own axis, (b) adjust the yaw angle of the right-angled probe tip, and (c) revolve the probe about the axis common to the positioner and casing. A cam plate engages a cam-follower portion of the probe and normally rotates with the positioner. The positioner includes a first-motor-driven ring gear which effects slidable movement of the probe by rotating the positioner at a time when an external pneumatic cylinder is actuated to engage the cam plate and hold it stationary. When the pneumatic cylinder is not actuated, this ring gear can be driven to revolve the positioner and thus the probe to a desired circumferential location about the above-mentioned common axis. A second motor-driven ring gear included in the positioner can be driven to rotate the probe about its axis, thus adjusting the yaw angle of the probe tip. The positioner can be used in highly corrosive atmosphere, such as gaseous uranium hexafluoride. 10 claims, 6 figures

Influence of probe geometry on the response of an electrostatic probe

DEFF Research Database (Denmark)

Johansson, Torben; Crichton, George C; McAllister, Iain Wilson

1999-01-01

The response of an electrostatic probe is examined with reference to the probe geometry. The study involves the evaluation of the probe lambda function, from which response-related characteristic parameters can be derived. These parameters enable the probe detection sensitivity Se and spatial...

Anti-theft device staining on banknotes detected by mass spectrometry imaging.

Science.gov (United States)

Correa, Deleon Nascimento; Zacca, Jorge Jardim; Rocha, Werickson Fortunato de Carvalho; Borges, Rodrigo; de Souza, Wanderley; Augusti, Rodinei; Eberlin, Marcos Nogueira; Vendramini, Pedro Henrique

2016-03-01

We describe the identification and limits of detection of ink staining by mass spectrometry imaging (MSI), as used in anti-theft devices (ATDs). Such ink staining is applied to banknotes during automated teller machine (ATM) explosions. Desorption electrospray ionization (DESI) coupled with high-resolution and high-accuracy orbitrap mass spectrometry (MS) and a moving stage device were applied to obtain 2D molecular images of the major dyes used for staining, that is, 1-methylaminoanthraquinone (MAAQ), rhodamine B (RB) and rhodamine 6G (R6G). MAAQ could not be detected because of its inefficient desorption by DESI from the banknote cellulose surface. By contrast, ATD staining on banknotes is perceptible by the human naked eye only at concentrations higher than 0.2 μg cm(-2), whereas both RB and R6G at concentrations 200 times lower (as low as 0.001 μg cm(-2)) could be easily detected and imaged by DESI-MSI, with selective and specific identification of each analyte and their spatial distribution on samples from suspects. This technique is non-destructive, and no sample preparation is required, which ensures sample preservation for further forensic investigations. Copyright © 2016. Published by Elsevier Ireland Ltd.

Effectiveness of Vascular Markers (Immunohistochemical Stains) in Soft Tissue Sarcomas.

Science.gov (United States)

Naeem, Namra; Mushtaq, Sajid; Akhter, Noreen; Hussain, Mudassar; Hassan, Usman

2018-05-01

To ascertain the effectiveness of IHC markers of vascular origin like CD31, CD34, FLI1 and ERG in vascular soft tissue sarcomas including angiosarcomas, Kaposi sarcomas, epithelioid hemangioendothelioma and a non-vascular soft tissue sarcoma (Epithelioid sarcoma). Descriptive study. Shaukat Khanum Memorial Cancer Hospital and Research Centre, Lahore, from 2011 to 2017. Diagnosed cases of angiosarcomas (n=48), epithelioid hemangioendothelioma (n=9), Kaposi sarcoma (n=9) and epithelioid sarcoma (n=20) were selected. Immunohistochemical staining as performed on formalin fixed paraffin embedded sections. The sections were stained for the following markers: CD34 (VENTANA clone Q Bend 10), CD31 (Leica clone 1 A 10), FLI1 (CELL MARQUE clone MRQ-1) and ERG (CELL MARQUE clone EP111). A complete panel of CD34, CD31 and ERG was applied on 8/48 cases of angiosarcomas with triple positivity in 6 cases. Eight cases showed positivity for only CD31 and ERG and 2 cases showed positivity for only ERG. A complete panel of CD34, CD31 and ERG was applied on 3/9 cases of epithelioid hemangioendothelioma with positivity for all markers in 2 cases. Combined positivity for ERG and CD34 was seen in 2 cases and on 4 cases only CD31 immunohistochemical was solely applied with 100% positivity. FLI1 was not applied on any case. Among 9 cases of Kaposi sarcoma, ERG, CD34 and CD31 in combination were applied on only 1 case with triple positivity. Remaining cases show positivity for either CD34, CD31 or FLI1. Majority of cases of epithelioid sarcomas were diagnosed on the basis of cytokeratin and CD34 positivity with loss of INI1. The other vascular markers showed negativity in all cases. Among these four markers, ERG immunohistochemical stain is highly effective for endothelial differentiation due to its specific nuclear staining pattern in normal blood vessel endothelial cells (internal control) as well as neoplastic cells of vascular tumors and lack of background staining.

Porcine intestinal mast cells. Evaluation of different fixatives for histochemical staining techniques considering tissue shrinkage

Directory of Open Access Journals (Sweden)

J. Rieger

2013-07-01

Full Text Available Staining of mast cells (MCs, including porcine ones, is critically dependent upon the fixation and staining technique. In the pig, mucosal and submucosal MCs do not stain or stain only faintly after formalin fixation. Some fixation methods are particularly recommended for MC staining, for example the fixation with Carnoy or lead salts. Zinc salt fixation (ZSF has been reported to work excellently for the preservation of fixation-sensitive antigens. The aim of this study was to establish a reliable histological method for counting of MCs in the porcine intestinum. For this purpose, different tissue fixation and staining methods that also allow potential subsequent immunohistochemical investigations were evaluated in the porcine mucosa, as well as submucosa of small and large intestine. Tissues were fixed in Carnoy, lead acetate, lead nitrate, Zamboni and ZSF and stained subsequently with either polychromatic methylene blue, alcian blue or toluidine blue. For the first time our study reveals that ZSF, a heavy metal fixative, preserves metachromatic staining of porcine MCs. Zamboni fixation was not suitable for histochemical visualization of MCs in the pig intestine. All other tested fixatives were suitable. Alcian blue and toluidine blue co-stained intestinal goblet cells which made a prima facie identification of MCs difficult. The polychromatic methylene blue proved to be the optimal staining. In order to compare MC counting results of the different fixation methods, tissue shrinkage was taken into account. As even the same fixation caused shrinkage-differences between tissue from small and large intestine, different factors for each single fixation and intestinal localization had to be calculated. Tissue shrinkage varied between 19% and 57%, the highest tissue shrinkage was found after fixation with ZSF in the large intestine, the lowest one in the small intestine after lead acetate fixation. Our study emphasizes that MC counting results from

The Oxford Probe: an open access five-hole probe for aerodynamic measurements

Science.gov (United States)

Hall, B. F.; Povey, T.

2017-03-01

The Oxford Probe is an open access five-hole probe designed for experimental aerodynamic measurements. The open access probe can be manufactured by the end user via additive manufacturing (metal or plastic). The probe geometry, drawings, calibration maps, and software are available under a creative commons license. The purpose is to widen access to aerodynamic measurement techniques in education and research environments. There are many situations in which the open access probe will allow results of comparable accuracy to a well-calibrated commercial probe. We discuss the applications and limitations of the probe, and compare the calibration maps for 16 probes manufactured in different materials and at different scales, but with the same geometrical design.

The Oxford Probe: an open access five-hole probe for aerodynamic measurements

International Nuclear Information System (INIS)

Hall, B F; Povey, T

2017-01-01

The Oxford Probe is an open access five-hole probe designed for experimental aerodynamic measurements. The open access probe can be manufactured by the end user via additive manufacturing (metal or plastic). The probe geometry, drawings, calibration maps, and software are available under a creative commons license. The purpose is to widen access to aerodynamic measurement techniques in education and research environments. There are many situations in which the open access probe will allow results of comparable accuracy to a well-calibrated commercial probe. We discuss the applications and limitations of the probe, and compare the calibration maps for 16 probes manufactured in different materials and at different scales, but with the same geometrical design. (paper)

Supravital dithizone staining in the isolation of human and rat pancreatic islets

DEFF Research Database (Denmark)

Hansen, W A; Christie, M R; Kahn, R

1989-01-01

Dithizone, a zinc chelating agent, is known to selectively stain the islets of Langerhans in the pancreas. In the present study, we have used this stain to aid the identification of islets in material obtained by collagenase digestion of human pancreas. Islets were shown to rapidly and reversibly...... techniques for the large scale isolation of functionally intact human islets....

a comparison of modified and standard papanicolaou staining ...

African Journals Online (AJOL)

2011-07-07

Jul 7, 2011 ... modified pap method and standard Papanicolaou method respectively. The staining characteristics in .... alcohol was replaced by 0.5 % acetic acid and also, .... was 37.1, standard deviation of 8.0 and a median of. 36.5 years.

Probe Techniques. Introductory Remarks

Energy Technology Data Exchange (ETDEWEB)

Emeleus, K. G. [School of Physics and Applied Mathematics, Queen' s University, Belfast (United Kingdom)

1968-04-15

In this brief introduction to the session on probes, the history of theii development is first touched on briefly. Reference is then made to the significance of the work to be described by Medicus, for conductivity and recombination calculations, and by Lam and Su, for a wide range of medium and higher pressure plasmas. Finally, a number of other probe topics are mentioned, including multiple probes; probes in electronegative plasmas; resonance probes; probes in noisy discharges; probes as oscillation detectors; use of probes where space-charge is not negligible. (author)

Modified Cajal's trichrome stain as a diagnostic aid in the study of epithelial pathology

Directory of Open Access Journals (Sweden)

Karpagaselvi Sanjai

2017-01-01

Full Text Available Background: Diagnosis of initial epithelial pathology maybe difficult in Squamous Cell Carcinoma (SCC, Carcinoma In Situ and other atypical epithelial malignancies, under routine Haematoxylin and Eosin (H and E stain. The detection of minor basement membrane alterations in doubtful cases is both time consuming and confusing. Aims: To evaluate efficacy of Modified Cajal's Trichrome Stain (CTS in relation to Haematoxylin and Eosin for study of epithelial dysplasia, carcinoma in situ, micro invasive SCC, frank SCC, and SCC in lymph nodes. Materials and Methods: Formalin-fixed, paraffin-embedded tissue blocks of mild epithelial dysplasia (n = 2, moderate epithelial dysplasia (n = 2, severe epithelial dysplasia (n = 4, carcinoma in situ (n = 1, micro-invasive SCC (n = 4, verrucous carcinoma (n = 1, and frank OSCC (n = 5 were stained with CTS and H&E. The sections were compared based on set histopathological criteria. Results and Conclusion: In SCC cases stained with CTS, invasion into connective tissue and keratin pearls were strikingly evident. Depth of invasion could be more accurately determined. Tumour cells in lymph node were intensely contrasted and easily discernible. Thus, CTS is a good differential stain, clearly delineating the epithelial elements from the connective tissue elements visually. This helps in tracing the basement membrane very clearly. It is an economic, rapid and easy to use method which cannot replace Haematoxylin and Eosin stain in cancer diagnosis, but can definitely be used adjunctive to it. Prompt diagnosis is crucial to effective treatment, and this stain assists in early and rapid diagnosis of cancer.

Probe-diverse ptychography

Energy Technology Data Exchange (ETDEWEB)

Peterson, I., E-mail: isaac.russellpeterson@rmit.edu.au [ARC Centre of Excellence for Coherent X-ray Science, the University of Melbourne, School of Physics, Victoria 3010 (Australia); Harder, R. [Advanced Photon Source, Argonne National Laboratory, Argonne, IL 60439 (United States); Robinson, I.K. [Research Complex at Harwell, Didcot, Oxfordshire OX11 0DE (United Kingdom); London Centre for Nanotechnology, University College London, London WC1H 0AH (United Kingdom)

2016-12-15

We propose an extension of ptychography where the target sample is scanned separately through several probes with distinct amplitude and phase profiles and a diffraction image is recorded for each probe and each sample translation. The resulting probe-diverse dataset is used to iteratively retrieve high-resolution images of the sample and all probes simultaneously. The method is shown to yield significant improvement in the reconstructed sample image compared to the image obtained using the standard single-probe ptychographic phase-retrieval scheme.

The application of anti-ESAT-6 monoclonal antibody fluorescent probe in ex vivo near-infrared fluorescence imaging in mice with pulmonary tuberculosis.

Science.gov (United States)

Feng, Feng; Zhang, Haoling; Zhu, Zhaoqin; Li, Cong; Shi, Yuxin; Zhang, Zhiyong

2014-09-01

Here, we aimed to assess the feasibility of anti-ESAT-6 monoclonal antibody (mAb) coupling with IR783 and rhodamine fluorescent probe in the detection of ESAT-6 expression in tuberculosis tissue of mice using near-infrared fluorescence imaging. IR783 and rhodamine were conjugated to the anti-ESAT-6 mAb or IgG. Mice in the experimental group were injected with fluorescence-labeled mAb probe, and mice in the control group were injected with fluorescence-labeled non-specific IgG antibody. Twenty-four hours later, the lung tissue of mice was examined using ex vivo near-infrared fluorescence imaging. In addition, the contrast-to-noise ratio (CNR) was calculated by measuring the signal intensities of the pulmonary lesions, normal lung tissue and background noise. The frozen lung tissue section was examined under fluorescence microscopy and compared with hemoxylin and eosin (HE) staining. The ex vivo near-infrared fluorescence imaging showed that the fluorescence signal in the lung tuberculosis lesions in the experimental group was significantly enhanced, whereas there was only a weak fluorescence signal or even no fluorescence signal in the control group. CNR values were 64.40 ± 7.02 (n = 6) and 8.75 ± 3.87 (n = 6), respectively (t = 17.01, p fluorescence accumulation distribution detected under fluorescence microscopy was consistent with HE staining of the tuberculosis region. In conclusion, anti-ESAT-6 mAb fluorescent probe could target and be applied in specific ex vivo imaging of mice tuberculosis, and may be of further use in tuberculosis in living mice. Copyright © 2013 John Wiley & Sons, Ltd.

Cell wall staining with Trypan blue enables quantitative analysis of morphological changes in yeast cells

DEFF Research Database (Denmark)

Liesche, Johannes; Marek, Magdalena; Günther-Pomorski, Thomas

2015-01-01

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......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...... 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...

PERSEPSI DAN TRANSFORMASI VISI DAN MISI PADA CIVITAS AKADEMIKA STAIN PEKALONGAN

Directory of Open Access Journals (Sweden)

Imam Khanafi

2013-05-01

Full Text Available Penelitian ini mengukur tingkat persepsi visi dan misi civitas akademika STAIN Pekalongan, dan mengukur juga tingkat implementasi sebagai transformasi visi misi tersebut serta menemukan hubungan antara persepsi dengan implementasinya. Kajian ini memiliki signifikan yang strategis dalam konteks manajemen dan pengembangan Pendidikan di STAIN Pekalongan dan lembaga pendidikan lainnya. Hal ini karena posisi visi dan misi sebagai kerangka acuan (frame of reference yang mengarahkan dan menjelaskan kepada keseluruh unsur civitas akademika dalam setiap program kerjanya.

Teeth re-whitening effect of strawberry juice on coffee stained teeth

OpenAIRE

Annisya Pramesti; Tadeus Arufan Jasrin; Opik Taofik Hidayat

2018-01-01

Many people favor coffee. However, regarding health and aesthetic dentistry, coffee gives a negative effect. Tanin in coffee causes a brown stain on the tooth surface. Therefore, in aesthetic dental care, teeth whitening has become popular matter. One of the natural ingredients used for teeth whitening treatment is strawberry. The purpose of this study was to obtained data regarding the effect of strawberry juice on the re-whitening process of the coffee-stained tooth enamel surface. This stu...

INTERNALISASI PENDIDIKAN KARAKTER DI PERGURUAN TINGGI: Studi Kasus di Jurusan Tarbiyah STAIN PONOROGO

Directory of Open Access Journals (Sweden)

Kharisul Wathoni

2016-05-01

Full Text Available This study will examine the internalization of character values at the Tarbiyah Department of STAIN Ponorogo. It has been found out that STAIN Ponorogo, in particular the Tarbiyah Department, has made efforts to undertake character education to the students through three patterns: during the learning process, during process of academic administrative services, and during extracurricular and intraccurricular activities followed by students at STAIN Ponorogo. The characters to be internalized are honesty, discipline, religiousity, creativity, self-reliance, responsibility, tolerance, communication and responsibility. Keywords: character education, morality, internalization

Electrical resistivity probes

Science.gov (United States)

Lee, Ki Ha; Becker, Alex; Faybishenko, Boris A.; Solbau, Ray D.

2003-10-21

A miniaturized electrical resistivity (ER) probe based on a known current-voltage (I-V) electrode structure, the Wenner array, is designed for local (point) measurement. A pair of voltage measuring electrodes are positioned between a pair of current carrying electrodes. The electrodes are typically about 1 cm long, separated by 1 cm, so the probe is only about 1 inch long. The electrodes are mounted to a rigid tube with electrical wires in the tube and a sand bag may be placed around the electrodes to protect the electrodes. The probes can be positioned in a borehole or on the surface. The electrodes make contact with the surrounding medium. In a dual mode system, individual probes of a plurality of spaced probes can be used to measure local resistance, i.e. point measurements, but the system can select different probes to make interval measurements between probes and between boreholes.

Liquid dish washing soap: An excellent substitute for xylene and alcohol in hematoxylin and eosin staining procedure

Directory of Open Access Journals (Sweden)

Surekha Ramulu

2012-01-01

Full Text Available Aims: Liquid dish washing solution (DWS was used as a substitute for xylene to dewax tissue sections during hematoxylin and eosin (H and E staining. The aim was to test and compare the hypothesis that xylene-ethanol free (XEF sections deparaffinized with diluted DWS are better than or at par with the conventional H and E sections. Materials and Methods: Fifty paraffin-embedded tissue blocks was included. One section was stained with conventional HandE (group A and the other with XEF HandE (group B staining method. Slides were scored for parameters: nuclear, cytoplasmic, clarity, uniformity, and crispness of staining. Z test was used for statistical analysis. For accuracy of diagnosis, sensitivity, specificity, positive predictive value, and negative predictive value were tested. Results: Adequate nuclear staining was noted in 94% in group A and 96% in group B, -adequate cytoplasmic staining in 92% in group A and 86% in group B, clarity in 94% of group A and 96% of group B sections, uniform staining in 92% of group A and 80% of group B sections, crisp stain in 96% of group A and 88% of group B sections, and 94% of group A sections stained adequately for diagnosis as compared with 90% in group B sections. Conclusion: Liquid DWS can be used as an alternative and effective substitute to xylene and ethanol in routine HandE staining procedure.

Nonlinear multicontrast microscopy of hematoxylin-and-eosin-stained histological sections

Science.gov (United States)

Tuer, Adam; Tokarz, Danielle; Prent, Nicole; Cisek, Richard; Alami, Jennifer; Dumont, Daniel J.; Bakueva, Ludmila; Rowlands, John; Barzda, Virginijus

2010-03-01

Imaging hematoxylin-and-eosin-stained cancerous histological sections with multicontrast nonlinear excitation fluorescence, second- and third-harmonic generation (THG) microscopy reveals cellular structures with extremely high image contrast. Absorption and fluorescence spectroscopy together with second hyperpolarizability measurements of the dyes shows that strong THG appears due to neutral hemalum aggregation and is subsequently enhanced by interaction with eosin. Additionally, fluorescence lifetime imaging microscopy reveals eosin fluorescence quenching by hemalums, showing better suitability of only eosin staining for fluorescence microscopy. Multicontrast nonlinear microscopy has the potential to differentiate between cancerous and healthy tissue at a single cell level.

Lack of clinical utility of urine gram stain for suspected urinary tract infection in pediatric patients.

Science.gov (United States)

Cantey, Joseph B; Gaviria-Agudelo, Claudia; McElvania TeKippe, Erin; Doern, Christopher D

2015-04-01

Urinary tract infection (UTI) is one of the most common infections in children. Urine culture remains the gold standard for diagnosis, but the utility of urine Gram stain relative to urinalysis (UA) is unclear. We reviewed 312 pediatric patients with suspected UTI who had urine culture, UA, and urine Gram stain performed from a single urine specimen. UA was considered positive if ≥10 leukocytes per oil immersion field were seen or if either nitrates or leukocyte esterase testing was positive. Urine Gram stain was considered positive if any organisms were seen. Sensitivity, specificity, and positive and negative predictive values were calculated using urine culture as the gold standard. Thirty-seven (12%) patients had a culture-proven UTI. Compared to urine Gram stain, UA had equal sensitivity (97.3% versus 97.5%) and higher specificity (85% versus 74%). Empirical therapy was prescribed before the Gram stain result was known in 40 (49%) patients and after in 42 (51%) patients. The antibiotics chosen did not differ between the two groups (P=0.81), nor did they differ for patients with Gram-negative rods on urine Gram stain compared to those with Gram-positive cocci (P=0.67). From these data, we conclude that UA has excellent negative predictive value that is not enhanced by urine Gram stain and that antibiotic selection did not vary based on the urine Gram stain result. In conclusion, the clinical utility of urine Gram stain does not warrant the time or cost it requires. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Development of species-specific rDNA probes for Giardia by multiple fluorescent in situ hybridization combined with immunocytochemical identification of cyst wall antigens.

Science.gov (United States)

Erlandsen, Stanley L; Jarroll, Edward; Wallis, Peter; van Keulen, Harry

2005-08-01

In this study, we describe the development of fluorescent oligonucleotide probes to variable regions in the small subunit of 16S rRNA in three distinct Giardia species. Sense and antisense probes (17-22 mer) to variable regions 1, 3, and 8 were labeled with digoxygenin or selected fluorochomes (FluorX, Cy3, or Cy5). Optimal results were obtained with fluorochome-labeled oligonucleotides for detection of rRNA in Giardia cysts. Specificity of fluorescent in situ hybridization (FISH) was shown using RNase digestion and high stringency to diminish the hybridization signal, and oligonucleotide probes for rRNA in Giardia lamblia, Giardia muris, and Giardia ardeae were shown to specifically stain rRNA only within cysts or trophozoites of those species. The fluorescent oligonucleotide specific for rRNA in human isolates of Giardia was positive for ten different strains. A method for simultaneous FISH detection of cysts using fluorescent antibody (genotype marker) and two oligonucleotide probes (species marker) permitted visualization of G. lamblia and G. muris cysts in the same preparation. Testing of an environmental water sample revealed the presence of FISH-positive G. lamblia cysts with a specific rDNA probe for rRNA, while negative cysts were presumed to be of animal or bird origin.

Laser Treatment of Port Wine Stains

Science.gov (United States)

Majaron, Boris; Nelson, J. Stuart

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

Lectins stain cells differentially in the coral, Montipora capitata

Science.gov (United States)

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.

Acridine orange staining and radiometric detection of microorganisms in blood cultures

International Nuclear Information System (INIS)

Burdash, N.M.; Manos, J.P.; Bannister, E.R.; Welborn, A.L.

1983-01-01

To determine whether acridine orange (AO) staining of blood cultures could be used as a substitute for blind subculture when used in conjunction with the BACTEC system (Johnston Laboratories, Inc., Towson, Md.), the two methods were compared on all BACTEC-negative specimens. Since blind subcultures were routinely performed in our laboratory on days 2 and 6 of incubation, AO staining was also performed on these days. Cultures which were BACTEC positive on day 1 of incubation were not included in the study. Of the 2,395 bottles tested after 2 days of incubation, 106 were subculture positive. Of these, 96 (90.6%) were also AO positive and BACTEC positive, 3 (2.8%) were AO positive and BACTEC negative, and 7 (6.6%) were AO negative and BACTEC positive. Of the 3,487 bottles tested on day 6 of incubation, 14 were subculture positive; 7 (50%) of these were AO positive and BACTEC positive, and seven were AO positive and BACTEC negative. Of the total of 10 culture-positive bottles missed by BACTEC, all were positive, and all 10 companion aerobic bottles were BACTEC positive. In both phases of the experiment, there was a total of only four false-positive AO stains. As a result of this investigation, we have substituted AO staining for blind subculturing of BACTEC-negative bottles

Evaluation of Staining-Dependent Colour Changes in Resin Composites Using Principal Component Analysis.

Science.gov (United States)

Manojlovic, D; Lenhardt, L; Milićević, B; Antonov, M; Miletic, V; Dramićanin, M D

2015-10-09

Colour changes in Gradia Direct™ composite after immersion in tea, coffee, red wine, Coca-Cola, Colgate mouthwash, and distilled water were evaluated using principal component analysis (PCA) and the CIELAB colour coordinates. The reflection spectra of the composites were used as input data for the PCA. The output data (scores and loadings) provided information about the magnitude and origin of the surface reflection changes after exposure to the staining solutions. The reflection spectra of the stained samples generally exhibited lower reflection in the blue spectral range, which was manifested in the lower content of the blue shade for the samples. Both analyses demonstrated the high staining abilities of tea, coffee, and red wine, which produced total colour changes of 4.31, 6.61, and 6.22, respectively, according to the CIELAB analysis. PCA revealed subtle changes in the reflection spectra of composites immersed in Coca-Cola, demonstrating Coca-Cola's ability to stain the composite to a small degree.

Laser beam diameter for port wine stain treatment

NARCIS (Netherlands)

Keijzer, M.; Pickering, J. W.; van Gemert, M. J.

1991-01-01

Optimal port wine stain treatment requires the selective absorption of light by the ectatic blood vessels. We investigated whether deeper blood vessels can be coagulated, without damaging other cutaneous structures, by varying the laser beam diameter. The penetration of the light was simulated with

Cell wall staining with Trypan blue enables quantitative analysis of morphological changes in yeast cells.

Science.gov (United States)

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.

Cell wall staining with Trypan Blue enables quantitative analysis of morphological changes in yeast cells

Directory of Open Access Journals (Sweden)

Johannes eLiesche

2015-02-01

Full Text Available 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.

Hematoxylin and eosin stain shows a high sensitivity but sub-optimal specificity in demonstrating iron pigment in liver biopsies.

Science.gov (United States)

Alwahaibi, Nasar Yousuf; Alkhatri, Azza Sarhan; Kumar, Johanes Selva

2015-01-01

Perls' stain is routinely used to demonstrate iron in liver biopsies. We tested the hypothesis that it may be unnecessary in cases, where no iron or another similar pigment was seen on the routine hematoxylin and eosin (H and E) stained section. The aim of this study was to evaluate the efficiency of H and E stain in demonstrating iron in liver biopsies as well as to determine the possibility of replacing Perls' stain with H and E stain. Two hundred pairs of slides of liver biopsies were taken from the archival files of the Department of Pathology from 2006 to 2011. Perls' and H and E slides were independently reviewed for the presence of iron. Hundred and one cases showed the presence of iron using H and E stain. 84 of 86 cases showed positive iron using both Perls' and H and E stains. Seventeen cases were positive using H and E stain but negative with Perls'. Only two cases did not show the presence of iron using H and E stain. Ninety-seven cases were negative using both Perls' and H and E stains. H and E stain showed a sensitivity, specificity, accuracy, positive predictive valve, and negative predictive value of 97.67%, 85.08%, 90.5%, 83.16%, and 97.98%, respectively. We demonstrate that the H and E stain is a sensitive method to detect iron pigment in liver biopsies, particularly when present in large quantities. A negative H and E stain might obviate the need for extra Perls' staining, thus saving costs and shortening report turn-around times.

Flow cytometric detection of micronuclei by combined staining of DNA and membranes

International Nuclear Information System (INIS)

Wessels, J.M.; Nuesse, M.

1995-01-01

A new staining method is presented for flow cytometric measurement of micronuclei (MN) in cell cultures and human lymphocytes using membrane-specific fluorescent dyes in addition to DNA staining. Several combinations of fluorescent membrane and DNA dyes were studied for a better discrimination of MN from debris in a suspension of nuclei and micronuclei. For staining of membranes, the lipophilic dyes 2-hydroxyethyl-7,12,17-tris(methoxyethyl)porphycene (HEPn) and 1,6-diphenyl-1,3,5-hexatriene (DPH) were used in combination with ethidium bromide (EB), proflavine (PF), and Hoechst 33258 (HO). Due to their spectral properties, HO or EB combined with HEPn were not as suitable for the discrimination of MN from debris as was HEPn in combination with PF. With HEPn in combination with PF, however, additional noise was found at low fluorescence intensities, probably due to free fluorescent dye molecules in the solution. The optimal simultaneous staining of membranes and DNA was obtained using a combination of DPH and EB. The induction of MN in Chinese hamster and mouse NIH-3T3 cells by UV-B illumination was studied with this new staining technique. UV-B illumination (280-360 nm) induced MN in both cell lines. Chinese hamster cells were found to be more sensitive to these wavelengths. Illumination with wavelengths above 360 nm did not induce MN in either cell line. The results obtained from human lymphocytes using the combination of EB or DPH were comparable to the results obtained with the combination of EB and HO. 23 refs., 7 figs

Staining of E-selectin ligands on paraffin-embedded sections of tumor tissue.

Science.gov (United States)

Carrascal, Mylène A; Talina, Catarina; Borralho, Paula; Gonçalo Mineiro, A; Henriques, Ana Raquel; Pen, Cláudia; Martins, Manuela; Braga, Sofia; Sackstein, Robert; Videira, Paula A

2018-05-02

The E-selectin ligands expressed by cancer cells mediate adhesion of circulating cancer cells to endothelial cells, as well as within tissue microenvironments important for tumor progression and metastasis. The identification of E-selectin ligands within cancer tissue could yield new biomarkers for patient stratification and aid in identifying novel therapeutic targets. The determinants of selectin ligands consist of sialylated tetrasaccharides, the sialyl Lewis X and A (sLe X and sLe A ), displayed on protein or lipid scaffolds. Standardized procedures for immunohistochemistry make use of the antibodies against sLe X and/or sLe A . However, antibody binding does not define E-selectin binding activity. In this study, we developed an immunohistochemical staining technique, using E-selectin-human Ig Fc chimera (E-Ig) to characterize the expression and localization of E-selectin binding sites on paraffin-embedded sections of different cancer tissue. E-Ig successfully stained cancer cells with high specificity. The E-Ig staining show high reactivity scores in colon and lung adenocarcinoma and moderate reactivity in triple negative breast cancer. Compared with reactivity of antibody against sLe X/A , the E-Ig staining presented higher specificity to cancer tissue with better defined borders and less background. The E-Ig staining technique allows the qualitative and semi-quantitative analysis of E-selectin binding activity on cancer cells. The development of accurate techniques for detection of selectin ligands may contribute to better diagnostic and better understanding of the molecular basis of tumor progression and metastasis.

Cyclometalated Iridium(III) Complexes as AIE Phosphorescent Probes for Real-Time Monitoring of Mitophagy in Living Cells

Science.gov (United States)

Jin, Chengzhi; Liu, Jiangping; Chen, Yu; Guan, Ruilin; Ouyang, Cheng; Zhu, Yanjiao; Ji, Liangnian; Chao, Hui

2016-02-01

Mitophagy, which is a special autophagy that removes damaging mitochondria to maintain sufficient healthy mitochondria, provides an alternative path for addressing dysfunctional mitochondria and avoiding cellular death. In the present study, by coupling the triphenylamine group with 2-phenylimidazo[4,5-f][1,10]phenanthroline derivatives, we synthesized five Ir(III) complexes with an AIE property that are expected to fulfill requirements for real-time monitoring of mitophagy. Ir1-Ir5 were exploited to image mitochondria with a short incubation time by confocal microscopy and inductive coupled plasma-mass spectrometry (ICP-MS). Due to aggregation-induced emission (AIE), Ir1-Ir5 exhibited excellent photostability compared to MitoTracker Green (MTG). Moreover, Ir1-Ir5 manifested satisfactory photostability in the mitochondrial physiological pH range. In addition, the uptake mechanism of Ir1 was investigated using confocal microscopy and flow cytometry analysis. Finally, using both Ir1 and LysoTracker Green, we were able to achieve real-time monitoring of mitophagy.

High spatial resolution Kelvin probe force microscopy with coaxial probes

International Nuclear Information System (INIS)

Brown, Keith A; Westervelt, Robert M; Satzinger, Kevin J

2012-01-01

Kelvin probe force microscopy (KPFM) is a widely used technique to measure the local contact potential difference (CPD) between an AFM probe and the sample surface via the electrostatic force. The spatial resolution of KPFM is intrinsically limited by the long range of the electrostatic interaction, which includes contributions from the macroscopic cantilever and the conical tip. Here, we present coaxial AFM probes in which the cantilever and cone are shielded by a conducting shell, confining the tip–sample electrostatic interaction to a small region near the end of the tip. We have developed a technique to measure the true CPD despite the presence of the shell electrode. We find that the behavior of these probes agrees with an electrostatic model of the force, and we observe a factor of five improvement in spatial resolution relative to unshielded probes. Our discussion centers on KPFM, but the field confinement offered by these probes may improve any variant of electrostatic force microscopy. (paper)

Evaluation of biosafe alternatives as xylene substitutes in hematoxylin and eosin staining procedure: A comparative pilot study

Science.gov (United States)

Sravya, Taneeru; Rao, Guttikonda Venkateswara; Kumari, Masabattula Geetha; Sagar, Yerraguntla Vidya; Sivaranjani, Yeluri; Sudheerkanth, Kondamarri

2018-01-01

Background: Xylene is synthetic hydrocarbon produced from coal tar known for its wide usage as universal solvent which has many hazardous effects. The aim of this study is to compare the efficacy of xylene-free hematoxylin and eosin (H and E) sections with conventional H and E sections. Materials and Methods: The study included ninety paraffin-embedded tissue blocks. Of these, sixty blocks were processed with sesame oil (xylene alternative) and thirty blocks with xylene. The study sample was divided into three groups. Sixty sections which are taken from sesame oil-processed blocks were stained with xylene-free H and E staining method. In xylene-free staining method, 95% diluted lemon water (Group A) and 1.7% dish washing solution (DWS, Group B) were used as deparaffinizing agents whereas the remaining 30 sections were processed with xylene and stained with conventional H and E staining method (Group C). Slides were scored for the following parameters: (i) nuclear staining (adequate = score 1, inadequate = score 0), (ii) cytoplasmic staining (adequate = score 1, inadequate = score 0), (iii) uniformity (present = score 1, absent = score 0), (iv) clarity (present = score 1, absent = score 0) and (v) intensity (present = score 1, absent = score 0). Score ≤2 was considered inadequate for diagnosis while scores 3–5 were considered adequate for diagnosis. Results: Adequate nuclear staining was noted in 90% of sections of Group A and 100% each in Group B and Group C (P 0.05); adequate uniformity of staining in 53.3% of sections of Group A, 70% in Group B and 83.3% in Group C (P 0.05) and adequate intensity of staining in 76.7% sections of Group A, 93.3% in Group B and 100% in Group C (P < 0.05). Group C sections stained adequate for diagnosis (93.3%) followed by Group B (88.7%) and Group A (78%; P < 0.05). Conclusion: Tissues processed with sesame oil and stained using 1.7% DWS were found to be effective alternative to xylene.

Histological staining methods preparatory to laser capture microdissection significantly affect the integrity of the cellular RNA.

Science.gov (United States)

Wang, Hongyang; Owens, James D; Shih, Joanna H; Li, Ming-Chung; Bonner, Robert F; Mushinski, J Frederic

2006-04-27

Gene expression profiling by microarray analysis of cells enriched by laser capture microdissection (LCM) faces several technical challenges. Frozen sections yield higher quality RNA than paraffin-imbedded sections, but even with frozen sections, the staining methods used for histological identification of cells of interest could still damage the mRNA in the cells. To study the contribution of staining methods to degradation of results from gene expression profiling of LCM samples, we subjected pellets of the mouse plasma cell tumor cell line TEPC 1165 to direct RNA extraction and to parallel frozen sectioning for LCM and subsequent RNA extraction. We used microarray hybridization analysis to compare gene expression profiles of RNA from cell pellets with gene expression profiles of RNA from frozen sections that had been stained with hematoxylin and eosin (H&E), Nissl Stain (NS), and for immunofluorescence (IF) as well as with the plasma cell-revealing methyl green pyronin (MGP) stain. All RNAs were amplified with two rounds of T7-based in vitro transcription and analyzed by two-color expression analysis on 10-K cDNA microarrays. The MGP-stained samples showed the least introduction of mRNA loss, followed by H&E and immunofluorescence. Nissl staining was significantly more detrimental to gene expression profiles, presumably owing to an aqueous step in which RNA may have been damaged by endogenous or exogenous RNAases. RNA damage can occur during the staining steps preparatory to laser capture microdissection, with the consequence of loss of representation of certain genes in microarray hybridization analysis. Inclusion of RNAase inhibitor in aqueous staining solutions appears to be important in protecting RNA from loss of gene transcripts.

ENAMEL SUSCEPTIBILITY TO RED WINE STAINING AFTER 35% HYDROGEN PEROXIDE BLEACHING

Science.gov (United States)

Berger, Sandrine Bittencourt; Coelho, Alessandra Sanchez; Oliveira, Valéria Aparecida Pessatti; Cavalli, Vanessa; Giannini, Marcelo

2008-01-01

Concern has been expressed regarding the staining of enamel surface by different beverages after bleaching. This study investigated the influence of 35% hydrogen peroxide bleaching agents on enamel surface stained with wine after whitening treatments. Flat and polished bovine enamel surfaces were submitted to two commercially available 35% hydrogen peroxide bleaching agents or kept in 100% humidity, as a control group (n = 10). Specimens of all groups were immersed in red wine for 48 h at 37°C, immediately, 24 h or 1 week after treatments. All specimens were ground into powder and prepared for the spectrophotometric analysis. Data were subjected to two-way analysis of variance and Fisher's PLSD test at 5% significance level. The amount of wine pigments uptake by enamel submitted to bleaching treatments was statistically higher than that of control group, independently of the evaluation time. Results suggested that wine staining susceptibility was increased by bleaching treatments. PMID:19089218

Enamel susceptibility to red wine staining after 35% hydrogen peroxide bleaching

Directory of Open Access Journals (Sweden)

Sandrine Bittencourt Berger

2008-06-01

Full Text Available Concern has been expressed regarding the staining of enamel surface by different beverages after bleaching. This study investigated the influence of 35% hydrogen peroxide bleaching agents on enamel surface stained with wine after whitening treatments. Flat and polished bovine enamel surfaces were submitted to two commercially available 35% hydrogen peroxide bleaching agents or kept in 100% humidity, as a control group (n = 10. Specimens of all groups were immersed in red wine for 48 h at 37°C, immediately, 24 h or 1 week after treatments. All specimens were ground into powder and prepared for the spectrophotometric analysis. Data were subjected to two-way analysis of variance and Fisher's PLSD test at 5% significance level. The amount of wine pigments uptake by enamel submitted to bleaching treatments was statistically higher than that of control group, independently of the evaluation time. Results suggested that wine staining susceptibility was increased by bleaching treatments.

An Evaluation on the Importance of Phosphotungstic Acid Haematoxyiin (PTAH Staining in Differential Diagnosis of Salivary Gland Neoplasms

Directory of Open Access Journals (Sweden)

E Yazdi

2003-02-01

Full Text Available The differential diagnosis between pleomorphic adenoma, the most common salivary gland neoplasm, microscopically, and two other common salivary gland neoplasms, meaning adenoid cystic carcinoma and mucoepidermoid carcinoma, is difficut.The purpose of this study was to determine the differences between pleomorphic adenoma, adenoid cystic carcinoma and mucoepidermoid carcinoma regarding the staining intensity with PTAH and H&E and also to investigate the relationship between staining intensity with PTAH and malignancy grade in mucoepidermoid carcinoma. 72 paraffin embeded samples including 24 pleomorphic adenoma, 24 adenoid cystic carcinoma and 24 mucoepidermoid carcinoma were selected and stained with PTAH. The staining intensity in total, in nucleus and cytoplasm of tumoral cells were evaluated and compared with H&E staining, statistically. The results were analyzed by Kruskal-Wallis, and Wilcoxon signed ranks tests (P<0.05.The logistic model was presented to predict the degree of malignancies through the assessment of nucleus and cytoplasm staining intensity in tumoral cells, and the patient's age. In this study, a statistically significant relationship was observed between staining intensity by PTAH and H& E in nucleus of tumoral cells in pleomorphic adenoma. Moreover, statistically significant relation between staining intensity by PTAH and H & E in cytoplasm of tumoral cells in adenoid cystic carcinoma was found. But there was no relation in other cases. On the other hand, a statistically significant relation between intensity of staining in total, in nucleus and cytoplasm of tumoral cells and the type of tumor was found. No relation was obtained between malignancy grade of mucoepidermoid carcinoma and staining intensity in total, in nucleus and cytoplasm of tumoral cells. The presented logistic model indicated a direct relation between tumor malignancy with patient's age and staining intensity in nucleus of tumoral cells, but a

A comparision of modified and standard papanicolaou staining ...

African Journals Online (AJOL)

Objective: To compare modified and standard Papanicolaou (Pap) staining methods in the assessment of the cervical smears. Design: A descriptive cross sectional study. setting: Kenyatta National Hospital. Subjects: One hundred and sixty two women who were eligible for a pap smear and met the inclusion criteria.

Detergent-compatible proteases: microbial production, properties, and stain removal analysis.

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Niyonzima, Francois Niyongabo; More, Sunil

2015-01-01

Proteases are one of the most important commercial enzymes used in various industrial domains such as detergent and leather industries. The alkaline proteases as well as other detergent-compatible enzymes such as lipases and amylases serve now as the key components in detergent formulations. They break down various stains during fabric washing. The search for detergent-compatible proteases with better properties is a continuous exercise. The current trend is to use detergent-compatible proteases that are stable over a wide temperature range. Although the proteases showing stability at elevated pH have the capacity to be used in detergent formulations, their usage can be significant if they are also stable and compatible with detergent and detergent ingredients, and also able to remove protein stains. Despite the existence of some reviews on alkaline proteases, there is no specification for the use of alkaline proteases as detergent additives. The present review describes the detergent-compatible proteases tested as detergent additives. An overview was provided for screening, optimization, purification, and properties of detergent compatible proteases, with an emphasis on the stability and compatibility of the alkaline proteases with the detergent and detergent compounds, as well as stain removal examination methods.

Matrix Remodeling During Intervertebral Disc Growth and Degeneration Detected by Multichromatic FAST Staining

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Leung, Victor Y.L.; Chan, Wilson C.W.; Hung, Siu-Chun; Cheung, Kenneth M.C.; Chan, Danny

2009-01-01

Various imaging techniques have been used to assess degeneration of the intervertebral disc, including many histological methods, but cartilage-oriented histological stains do not clearly show the comparatively complex structures of the disc. In addition, there is no integrated method to assess efficiently both the compartmental organization and matrix composition in disc samples. In this study, a novel histological method, termed FAST staining, has been developed to investigate disc growth and degeneration by sequential staining with fast green, Alcian blue, Safranin-O, and tartrazine to generate multichromatic histological profiles (FAST profiles). This identifies the major compartments of the vertebra-disc region, including the cartilaginous endplate and multiple zones of the annulus fibrosus, by specific FAST profile patterns. A disc degeneration model in rabbit established using a previously described puncture method showed gradual but profound alteration of the FAST profile during disc degeneration, supporting continual alteration of glycosaminoglycan. Changes of the FAST profile pattern in the nucleus pulposus and annulus fibrosus of the postnatal mouse spine suggested matrix remodeling activity during the growth of intervertebral discs. In summary, we developed an effective staining method capable of defining intervertebral disc compartments in detail and showing matrix remodeling events within the disc. The FAST staining method may be used to develop a histopathological grading system to evaluate disc degeneration or malformation. (J Histochem Cytochem 57:249–256, 2009) PMID:19001641

Characteristics of weak base-induced vacuoles formed around individual acidic organelles.

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Hiruma, Hiromi; Kawakami, Tadashi

2011-01-01

We have previously found that the weak base 4-aminopyridine induces Brownian motion of acidic organelles around which vacuoles are formed, causing organelle traffic disorder in neurons. Our present study investigated the characteristics of vacuoles induced by weak bases (NH(4)Cl, aminopyridines, and chloroquine) using mouse cells. Individual vacuoles included acidic organelles identified by fluorescent protein expression. Mitochondria and actin filaments were extruded outside the vacuoles, composing the vacuole rim. Staining with amine-reactive fluorescence showed no protein/amino acid content in vacuoles. Thus, serous vacuolar contents are probably partitioned by viscous cytosol, other organelles, and cytoskeletons, but not membrane. The weak base (chloroquine) was immunochemically detected in intravacuolar organelles, but not in vacuoles. Early vacuolization was reversible, but long-term vacuolization caused cell death. The vacuolization and cell death were blocked by the vacuolar H(+)-ATPase inhibitor and Cl--free medium. Staining with LysoTracker or LysoSensor indicated that intravacuolar organelles were strongly acidic and vacuoles were slightly acidic. This suggests that vacuolization is caused by accumulation of weak base and H(+) in acidic organelles, driven by vacuolar H(+)-ATPase associated with Cl(-) entering, and probably by subsequent extrusion of H(+) and water from organelles to the surrounding cytoplasm.

[Comparison of the quick Gram stain method to the B&M modified and favor methods].

Science.gov (United States)

Osawa, Kayo; Kataoka, Nobumasa; Maruo, Toshio

2011-01-01

The Gram stain is an established method for bacterial identification, but the time needed to carry out this stain is 2-3 min. We attempted to shorten this time and stained a total of 70 clinical specimens isolated from using the Bartholomew & Mittwer (B&M) modified or Favor methods with a 3 s duration for washing and staining steps. Results were plotted and analyzed using a Hue Saturation Intensity (HSI) model. The range based on a plot of the two methods with the HSI model was presented as a reference interval. Our results indicated that 100% (35/35) of strains were Gram positive and 97.1% (34/35) were Gram negative for the quick B&M modified method. In the quick Favor method, 80.0% (28/35) were Gram positive and 68.6% (24/35) of strains were Gram negative. We propose that the quick B&M modified method is equivalent to the standard Gram staining method and is superior to the quick Favor method.

Silver-Stained Fibrin Zymography: Separation of Proteases and Activity Detection Using a Single Substrate-Containing Gel.

Science.gov (United States)

Park, Chang-Su; Kang, Dae-Ook; Choi, Nack-Shick

2017-01-01

Silver-stained fibrin zymography for separation of protease bands and activity detection using a single substrate gel was designed. The method takes advantage of the nano-scale sensitivity of both zymography and silver staining. After sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) in a gel containing fibrin (protease substrate), the gel was incubated in enzyme reaction buffer and the zymogram gel was silver-stained. Bands with protease activity were stained with silver in clear areas where the protein substrate had been degraded. The molecular sizes of proteases were accurately determined.

In-office bleaching efficacy on stain removal from CAD/CAM and direct resin composite materials.

Science.gov (United States)

Alharbi, Amal; Ardu, Stefano; Bortolotto, Tissiana; Krejci, Ivo

2018-01-01

To evaluate the efficacy of in-office bleaching on stain removal from stained resin composite and ceramic computer-assisted design/computer-assisted manufacturing (CAD/CAM) blocks and direct resin composites. Forty disk-shaped samples were fabricated from each of nine materials: six CAD/CAM (VITABLOCS Mark II, Paradigm MZ100, Exp Vita Hybrid Ceramic, VITA ENAMIC, Exp Kerr, and LAVA Ultimate) and three direct resin composites (Filtek Supreme, Venus Diamond, and Filtek Silorane). Samples were randomly divided into five groups (n = 8), each stained with a particular staining solution. Using a calibrated spectrophotometer and a black background, L*a*b* values were assessed before and after 120 days of staining. Samples were subjected to in-office bleaching using 40% hydrogen peroxide gel for one hour. At subsequent assessment, color change (ΔE) was calculated as the difference between L*a*b* values. Both ANOVA and the Duncan test were used to identify differences between groups (α = 0.05). Bleaching resulted in significant differences in ΔE values for all materials (P Bleaching efficacy was highly influenced by material composition and staining solution. Residual color values after bleaching for ceramic and hybrid ceramics ranged from -0.49 to 2.35, within the clinically acceptable maximum of 3.3. Values after bleaching for resin-based CAD/CAM ranged from -0.7 to 7.08 while direct resin composites values ranged from -1.47 to 25.13. Coffee left the greatest residual color on all materials. Based on material nature, 40% hydrogen peroxide bleaching can remove staining. The new resin-based CAD/CAM blocks showed promising results in terms of color stability. Bleaching using 40% hydrogen peroxide can be an effective method to remove stains from dental restorations. In this way, restoration replacement as a result of discoloration may no longer be necessary. © 2017 Wiley Periodicals, Inc.

Diagnostic performance of dual-staining cytology for cervical cancer screening: A systematic literature review.

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Tjalma, Wiebren A A

2017-03-01

Cervical cancer screening saves lives. Secondary prevention in cervical cancer screening relies on the results of primary cytology and/or HPV testing. However, primary screening with cytology has a low sensitivity, and HPV screening has a low specificity. This means that either cancers are missed, or women are over-treated. To improve performance outcomes, the concept of dual-stain cytology (CINtec ® PLUS Cytology test) has been introduced. In this approach, additional staining with p16/Ki-67 is performed in cases where cytology results are abnormal (LSIL or ASCUS) and/or HPV-positive. Another way to describe this approach might be "diagnostic" cytology. In order to assess the value of this "diagnostic cytology", a systematic literature review was conducted of dual-stain cytology performance across multiple studies until May 2016. In a Belgian screening population (women age 25-65 years), dual-stain cytology was significantly more sensitive (66%) and slightly less specific (-1.0%) than cytology. In the population referred to colposcopy or with abnormal cytology (ASCUS, LSIL), dual-staining showed a significantly higher increase in specificity, and a slightly lower sensitivity than HPV testing. Specificity gains resulted in fewer false positives and an increase in the number of correct referrals to colposcopy. Dual-staining with p16/Ki-67 cytology is an attractive biomarker approach for triage in cervical cancer screening. Copyright © 2017 Elsevier B.V. All rights reserved.

p16/Ki-67 Dual Stain Cytology for Detection of Cervical Precancer in HPV-Positive Women.

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Wentzensen, Nicolas; Fetterman, Barbara; Castle, Philip E; Schiffman, Mark; Wood, Shannon N; Stiemerling, Eric; Tokugawa, Diane; Bodelon, Clara; Poitras, Nancy; Lorey, Thomas; Kinney, Walter

2015-12-01

Human papillomavirus (HPV)-based cervical cancer screening requires triage markers to decide who should be referred to colposcopy. p16/Ki-67 dual stain cytology has been proposed as a biomarker for cervical precancers. We evaluated the dual stain in a large population of HPV-positive women. One thousand five hundred and nine HPV-positive women screened with HPV/cytology cotesting at Kaiser Permanente California were enrolled into a prospective observational study in 2012. Dual stain cytology was performed on residual Surepath material, and slides were evaluated for dual stain-positive cells. Disease endpoints were ascertained from the clinical database at KPNC. We evaluated the clinical performance of the assay among all HPV-positive women and among HPV-positive, cytology-negative women. We used internal benchmarks for clinical management to evaluate the clinical relevance of the dual stain assay. We evaluated sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the dual stain compared with Pap cytology. All statistical tests were two-sided. The dual stain had lower positivity (45.9%) compared with cytology at an ASC-US threshold (53.4%). For detection of CIN2+, the dual stain had similar sensitivity (83.4% vs 76.6%, P = .1), and statistically higher specificity (58.9% vs 49.6%, P < .001), PPV (21.0% vs 16.6%, P < .001), and NPV (96.4% vs 94.2%, P = .01) compared with cytology. Similar patterns were observed for CIN3+. Women with a positive test had high enough risk for referral to colposcopy, while the risk for women with negative tests was below a one-year return threshold based on current US management guidelines. Dual stain cytology showed good risk stratification for all HPV-positive women and for HPV-positive women with normal cytology. Additional follow-up is needed to determine how long dual stain negative women remain at low risk of precancer. Published by Oxford University Press 2015. This work is written by (a

Red alder kitchen cabinets—How does application of commercial stains influence customer choice?

Science.gov (United States)

David Nicholls; Joseph. Roos

2007-01-01

A better understanding of consumer reaction and preferences for red alder (Alnus rubra Bong.) secondary products will help Alaska producers in entering new markets. In this study, red alder kitchen cabinets were commercially stained to six different levels and displayed at home shows in Portland, Oregon, and Anchorage, Alaska. The stains simulated...

Staining pattern classification of antinuclear autoantibodies based on block segmentation in indirect immunofluorescence images.

Directory of Open Access Journals (Sweden)

Jiaqian Li

Full Text Available 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%.

Black stain and dental caries in Filipino schoolchildren.

NARCIS (Netherlands)

Heinrich-Weltzien, R.; Monse, B.; Palenstein Helderman, W.H. van

2009-01-01

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

Histological staining methods preparatory to laser capture microdissection significantly affect the integrity of the cellular RNA

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Li Ming-Chung

2006-04-01

Full Text Available Abstract Background Gene expression profiling by microarray analysis of cells enriched by laser capture microdissection (LCM faces several technical challenges. Frozen sections yield higher quality RNA than paraffin-imbedded sections, but even with frozen sections, the staining methods used for histological identification of cells of interest could still damage the mRNA in the cells. To study the contribution of staining methods to degradation of results from gene expression profiling of LCM samples, we subjected pellets of the mouse plasma cell tumor cell line TEPC 1165 to direct RNA extraction and to parallel frozen sectioning for LCM and subsequent RNA extraction. We used microarray hybridization analysis to compare gene expression profiles of RNA from cell pellets with gene expression profiles of RNA from frozen sections that had been stained with hematoxylin and eosin (H&E, Nissl Stain (NS, and for immunofluorescence (IF as well as with the plasma cell-revealing methyl green pyronin (MGP stain. All RNAs were amplified with two rounds of T7-based in vitro transcription and analyzed by two-color expression analysis on 10-K cDNA microarrays. Results The MGP-stained samples showed the least introduction of mRNA loss, followed by H&E and immunofluorescence. Nissl staining was significantly more detrimental to gene expression profiles, presumably owing to an aqueous step in which RNA may have been damaged by endogenous or exogenous RNAases. Conclusion RNA damage can occur during the staining steps preparatory to laser capture microdissection, with the consequence of loss of representation of certain genes in microarray hybridization analysis. Inclusion of RNAase inhibitor in aqueous staining solutions appears to be important in protecting RNA from loss of gene transcripts.

Blockface histology with optical coherence tomography: a comparison with Nissl staining.

Science.gov (United States)

Magnain, Caroline; Augustinack, Jean C; Reuter, Martin; Wachinger, Christian; Frosch, Matthew P; Ragan, Timothy; Akkin, Taner; Wedeen, Van J; Boas, David A; Fischl, Bruce

2014-01-01

Spectral domain optical coherence tomography (SD-OCT) is a high resolution imaging technique that generates excellent contrast based on intrinsic optical properties of the tissue, such as neurons and fibers. The SD-OCT data acquisition is performed directly on the tissue block, diminishing the need for cutting, mounting and staining. We utilized SD-OCT to visualize the laminar structure of the isocortex and compared cortical cytoarchitecture with the gold standard Nissl staining, both qualitatively and quantitatively. In histological processing, distortions routinely affect registration to the blockface image and prevent accurate 3D reconstruction of regions of tissue. We compared blockface registration to SD-OCT and Nissl, respectively, and found that SD-OCT-blockface registration was significantly more accurate than Nissl-blockface registration. Two independent observers manually labeled cortical laminae (e.g. III, IV and V) in SD-OCT images and Nissl stained sections. Our results show that OCT images exhibit sufficient contrast in the cortex to reliably differentiate the cortical layers. Furthermore, the modalities were compared with regard to cortical laminar organization and showed good agreement. Taken together, these SD-OCT results suggest that SD-OCT contains information comparable to standard histological stains such as Nissl in terms of distinguishing cortical layers and architectonic areas. Given these data, we propose that SD-OCT can be used to reliably generate 3D reconstructions of multiple cubic centimeters of cortex that can be used to accurately and semi-automatically perform standard histological analyses. © 2013.

A prospective study of the diagnostic utility of sputum Gram stain in pneumonia.

Science.gov (United States)

Anevlavis, Stavros; Petroglou, Niki; Tzavaras, Athanasios; Maltezos, Efstratios; Pneumatikos, Ioannis; Froudarakis, Marios; Anevlavis, Eleftherios; Bouros, Demosthenes

2009-08-01

Sputum Gram stain and culture have been said to be unreliable indicators of the microbiological diagnosis of bacterial pneumonia. The etiological diagnosis of pneumonia is surrounded by great degree of uncertainty. This uncertainty should be and can be calculated and incorporated in the diagnosis and treatment. To determine the diagnostic accuracy and diagnostic value of sputum Gram stain in etiological diagnosis and initial selection of antimicrobial therapy of bacterial community acquired pneumonia (CAP). DESIGN-METHOD: Prospective study of 1390 patients with CAP admitted January 2002-June 2008, to our institutions. Of the 1390 patients, 178 (12.8%) fulfilled the criteria for inclusion into this study (good-quality sputa and presence of the same microorganism in blood and sputum cultures which was used as gold standard for assessing the diagnostic accuracy and diagnostic value of sputum Gram stain). The sensitivity of sputum Gram stain was 0.82 for Pneumococcal pneumonia, 0.76 for Staphylococcal pneumonia, 0.79 for Haemophilus influenzae pneumonia and 0.78 for Gram-negative bacilli pneumonia. The specificity of sputum Gram stain was 0.93 for Pneumococcal pneumonia, 0.96 for Staphylococcal pneumonia, 0.96 for H. influenzae pneumonia and 0.95 for Gram-negative bacilli pneumonia. The positive likelihood ratio (LR+) was 11.58 for Pneumococcal pneumonia, 19.38 for Staphylococcal pneumonia, 16.84 for H. influenzae pneumonia, 14.26 for Gram-negative bacilli pneumonia. The negative likelihood ratio (LR-) was 0.20 for Pneumococcal pneumonia, 0.25 for Staphylococcal pneumonia, 0.22 for H. influenzae pneumonia, and 0.23 for Gram-negative bacilli pneumonia. Sputum Gram stain is a dependable diagnostic test for the early etiological diagnosis of bacterial CAP that helps in choosing orthological and appropriate initial antimicrobial therapy.

Colour stabilities of three types of orthodontic clear aligners exposed to staining agents.

Science.gov (United States)

Liu, Chen-Lu; Sun, Wen-Tian; Liao, Wen; Lu, Wen-Xin; Li, Qi-Wen; Jeong, Yunho; Liu, Jun; Zhao, Zhi-He

2016-12-16

The aim of this study was to evaluate and compare the colour stabilities of three types of orthodontic clear aligners exposed to staining agents in vitro. Sixty clear orthodontic aligners produced by three manufacturers (Invisalign, Angelalign, and Smartee) were immersed in three staining solutions (coffee, black tea, and red wine) and one control solution (distilled water). After 12-h and 7-day immersions, the aligners were washed in an ultrasonic cleaner and measured with a colourimeter. The colour changes (ΔE*) were calculated on the basis of the Commission Internationale de I'Eclairage L*a*b* colour system (CIE L*a*b*), and the results were then converted into National Bureau of Standards (NBS) units. Fourier transformation infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM) were conducted to observe the molecular and morphologic alterations to the aligner surfaces, respectively. The three types of aligners exhibited slight colour changes after 12 h of staining, with the exception of the Invisalign aligners stained with coffee. The Invisalign aligners exhibited significantly higher ΔE* values (ranging from 0.30 to 27.81) than those of the Angelalign and Smartee aligners (ΔE* values ranging from 0.33 to 1.89 and 0.32 to 1.61, respectively, Paligners did not exhibit significant chemical differences before and after the immersions. The SEM results revealed different surface alterations to the three types of aligner materials after the 7-day staining. The three types of aesthetic orthodontic appliances exhibited colour stability after the 12-h immersion, with the exception of the Invisalign aligners stained by coffee. The Invisalign aligners were more prone than the Angelalign and Smartee aligners to pigmentation. Aligner materials may be improved by considering aesthetic colour stability properties.

DNA probes

International Nuclear Information System (INIS)

Castelino, J.

1992-01-01

The creation of DNA probes for detection of specific nucleotide segments differs from ligand detection in that it is a chemical rather than an immunological reaction. Complementary DNA or RNA is used in place of the antibody and is labelled with 32 P. So far, DNA probes have been successfully employed in the diagnosis of inherited disorders, infectious diseases, and for identification of human oncogenes. The latest approach to the diagnosis of communicable and parasitic infections is based on the use of deoxyribonucleic acid (DNA) probes. The genetic information of all cells is encoded by DNA and DNA probe approach to identification of pathogens is unique because the focus of the method is the nucleic acid content of the organism rather than the products that the nucleic acid encodes. Since every properly classified species has some unique nucleotide sequences that distinguish it from every other species, each organism's genetic composition is in essence a finger print that can be used for its identification. In addition to this specificity, DNA probes offer other advantages in that pathogens may be identified directly in clinical specimens

EFFECT OF SILICATE ON GRAM STAINING AND VIABILITY OF PNEUMOCOCCI AND OTHER BACTERIA

Science.gov (United States)

MacLeod, Colin M.; Roe, Amy S.

1956-01-01

Application of silicate solutions to living or heat-killed pneumococci and to certain "viridans" streptococci causes their conversion from a Gram-positive to a Gram-negative state. The original staining properties can be restored by suspending the silicate-treated bacteria in alkaline solutions of various salts but not by simple washing in water. Living pneumococci and the strains of streptococci whose staining properties are similarly affected are killed when suspended in silicate solutions. In other Gram-positive species silicate causes conversion to Gram negativity but restoration to positivity occurs upon washing in water. In a third group of Gram-positive organisms silicate has no effect on the Gram reaction. The viability of organisms in these two groups is unaffected by silicate under the conditions employed. No effect on staining or viability of Gram-negative bacteria has been observed. The effects of silicate on staining and viability are inhibited by nutrient broth or whole serum but not by purified serum albumin. Lecithin, choline, and other substituted ammonium compounds also inhibit the effects of silicate on pneumococci. PMID:13306854

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

Science.gov (United States)

Yamada, Kengo; Yoshida, Katsuyuki; Sonoda, Ryoichi

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

A method for acetylcholinesterase staining of brain sections previously processed for receptor autoradiography.

Science.gov (United States)

Lim, M M; Hammock, E A D; Young, L J

2004-02-01

Receptor autoradiography using selective radiolabeled ligands allows visualization of brain receptor distribution and density on film. The resolution of specific brain regions on the film often can be difficult to discern owing to the general spread of the radioactive label and the lack of neuroanatomical landmarks on film. Receptor binding is a chemically harsh protocol that can render the tissue virtually unstainable by Nissl and other conventional stains used to delineate neuroanatomical boundaries of brain regions. We describe a method for acetylcholinesterase (AChE) staining of slides previously processed for receptor binding. AChE staining is a useful tool for delineating major brain nuclei and tracts. AChE staining on sections that have been processed for receptor autoradiography provides a direct comparison of brain regions for more precise neuroanatomical description. We report a detailed thiocholine protocol that is a modification of the Koelle-Friedenwald method to amplify the AChE signal in brain sections previously processed for autoradiography. We also describe several temporal and experimental factors that can affect the density and clarity of the AChE signal when using this protocol.

Quantitative gel electrophoresis: new records in precision by elaborated staining and detection protocols.

Science.gov (United States)

Deng, Xi; Schröder, Simone; Redweik, Sabine; Wätzig, Hermann

2011-06-01

Gel electrophoresis (GE) is a very common analytical technique for proteome research and protein analysis. Despite being developed decades ago, there is still a considerable need to improve its precision. Using the fluorescence of Colloidal Coomassie Blue -stained proteins in near-infrared (NIR), the major error source caused by the unpredictable background staining is strongly reduced. This result was generalized for various types of detectors. Since GE is a multi-step procedure, standardization of every single step is required. After detailed analysis of all steps, the staining and destaining were identified as the major source of the remaining variation. By employing standardized protocols, pooled percent relative standard deviations of 1.2-3.1% for band intensities were achieved for one-dimensional separations in repetitive experiments. The analysis of variance suggests that the same batch of staining solution should be used for gels of one experimental series to minimize day-to-day variation and to obtain high precision. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Metastatic hepatocellular carcinoma to the skin staining positive with HMB-45.

Science.gov (United States)

Gross, Joshua A; Perniciaro, Charles; Gross, David J; Barksdale, Sarah K

2012-02-01

Hepatocellular carcinoma (HCC) is uncommonly observed as a cutaneous metastasis. We report a 76-year-old man with metastatic HCC to the skin of the nasal ala, diagnosed antecedent to the primary tumor. HCC was confirmed by positive immunostaining with Hep Par 1 in tissue from the metastasis and from a needle biopsy of the primary lesion. In addition, tumor cells from both the metastasis and liver stained positive with HMB-45. To our knowledge, HMB-45 positive staining has not been reported in either primary or metastatic HCC.

Methylene Blue-Aided In Vivo Staining of Central Airways during Flexible Bronchoscopy

Directory of Open Access Journals (Sweden)

Sabine Zirlik

2012-01-01

Full Text Available Background. The early diagnosis of malignant and premalignant changes of the bronchial mucosa remains a major challenge during bronchoscopy. Intravital staining techniques are not new. Previous small case series suggested that analysis of the bronchial mucosal surface using chromoendoscopy allows a prediction between neoplastic and nonneoplastic lesions. Objectives. The aim of the present study was to evaluate chromobronchoscopy as a method to identify malignant and premalignant lesions in the central airways in a prospective manner. Methods. In 26 patients we performed chromoendoscopy with 0.1% methylene blue during ongoing flexible white light bronchoscopy. Circumscribed lesions in central airways were further analyzed by biopsies and histopathologic examination. Results. In the majority of cases neither flat nor polypoid lesions in the central airways were stained by methylene blue. In particular, exophytic growth of lung cancer did not show any specific pattern in chromobronchoscopy. However, a specific dye staining was detected in one case where exophytic growth of metastatic colorectal cancer was present in the right upper lobe. In two other cases, a circumscribed staining was noted in unsuspicious mucosa. But histology revealed inflammation only. Conclusions. In contrast to previous studies, the present findings clearly indicate that chromobronchoscopy is not useful for early detection of malignant or premalignant lesions of the central airways.

Counting probe

International Nuclear Information System (INIS)

Matsumoto, Haruya; Kaya, Nobuyuki; Yuasa, Kazuhiro; Hayashi, Tomoaki

1976-01-01

Electron counting method has been devised and experimented for the purpose of measuring electron temperature and density, the most fundamental quantities to represent plasma conditions. Electron counting is a method to count the electrons in plasma directly by equipping a probe with the secondary electron multiplier. It has three advantages of adjustable sensitivity, high sensitivity of the secondary electron multiplier, and directional property. Sensitivity adjustment is performed by changing the size of collecting hole (pin hole) on the incident front of the multiplier. The probe is usable as a direct reading thermometer of electron temperature because it requires to collect very small amount of electrons, thus it doesn't disturb the surrounding plasma, and the narrow sweep width of the probe voltage is enough. Therefore it can measure anisotropy more sensitively than a Langmuir probe, and it can be used for very low density plasma. Though many problems remain on anisotropy, computer simulation has been carried out. Also it is planned to provide a Helmholtz coil in the vacuum chamber to eliminate the effect of earth magnetic field. In practical experiments, the measurement with a Langmuir probe and an emission probe mounted to the movable structure, the comparison with the results obtained in reverse magnetic field by using a Helmholtz coil, and the measurement of ionic sound wave are scheduled. (Wakatsuki, Y.)

Trace element analysis of blackish staining on the crowns of human archaeological teeth

Energy Technology Data Exchange (ETDEWEB)

Stermer, E.M. [Univ. of Oslo, Dept. of Oral Radiology, Oslo (Norway); Risnes, S. [Univ. of Oslo, Dept. of Oral Biology, Faculty of Dentistry, Oslo (Norway); Fischer, P.M. [Chalmers Univ. of Technology, Dept. of Physics, SIMS Lab., Goeteborg (Sweden)

1996-10-01

A blackish staining found on the crowns of teeth of 51 skulls from the excavation of the medieval St. Olav`s church in Trondheim was analyzed using secondary ion mass spectrometry (SIMS) and atomic absorption spectrometry (AAS). In four teeth, mass spectra and step scan concentration profiles of SIMS were performed and compared with the grey scale pattern in photographs of the analyzed paths. The manganese curve showed the highest degree of conformity with the grey scale pattern. The AAS analysis confirmed the increased content of manganese in blackish stained enamel. It was concluded that manganese, probably in the form of an oxide deposited from the soil, was the cause of the blackish staining. (au).

Trace element analysis of blackish staining on the crowns of human archaeological teeth

International Nuclear Information System (INIS)

Stermer, E.M.; Risnes, S.; Fischer, P.M.

1996-01-01

A blackish staining found on the crowns of teeth of 51 skulls from the excavation of the medieval St. Olav's church in Trondheim was analyzed using secondary ion mass spectrometry (SIMS) and atomic absorption spectrometry (AAS). In four teeth, mass spectra and step scan concentration profiles of SIMS were performed and compared with the grey scale pattern in photographs of the analyzed paths. The manganese curve showed the highest degree of conformity with the grey scale pattern. The AAS analysis confirmed the increased content of manganese in blackish stained enamel. It was concluded that manganese, probably in the form of an oxide deposited from the soil, was the cause of the blackish staining. (au)

The Comparative Study of Art of Manufacturing Orosi and Stained Glass Windows in Iran and Europe

Directory of Open Access Journals (Sweden)

Zahra Sadat Abooei Mehrizi

2017-12-01

Full Text Available For a long time, glass manufacturing art has been globally common. There are certain similarities between Stained Glass and Orosi works. Based on historical texts, peak of Orosi art in Iran occurred during the reign of Safavid dynasty while its fall was after Qajar era. After introduction to churches after 12th century, Stained Glass manufacturing art was officially recognized. In contemporary era, Orosi art has almost faded away since it did not adapt to architecture. Similarly, Stained Glass art did not develop after it was introduced to Iran. The objectives of comparison between these arts are clarification of effects of the two arts on each other as well as better understanding of Orosi glass and Stained Glass arts. From methodological viewpoint, this study is descriptive-analytical in nature. In order to obtain better results, similar and available works of Orosi and Stained Glass arts were studied. The results of comparing these two arts suggested similar manufacturing methods and designs. In addition, a common application of the two arts is communication of greatness of the building to visitors’ minds. Stained Glass was mostly used for representation of Christ’s story while Orosi art was commonly used for depicting geometrical patterns. As a result, Orosi art was not initially influenced by Stained Glass art. In some cases, one could claim that Stained Glass art was influenced by Orosi art. Numerous factors played a role in creating differences between these two arts among which one could point to climatic conditions and patrons of such arts.

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

NARCIS (Netherlands)

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

2012-01-01

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

Utility of gram staining for evaluation of the quality of cystic fibrosis sputum samples.

Science.gov (United States)

Nair, Bindu; Stapp, Jenny; Stapp, Lynn; Bugni, Linda; Van Dalfsen, Jill; Burns, Jane L

2002-08-01

The microscopic examination of Gram-stained sputum specimens is very helpful in the evaluation of patients with community-acquired pneumonia and has also been recommended for use in cystic fibrosis (CF) patients. This study was undertaken to evaluate that recommendation. One hundred one sputum samples from CF patients were cultured for gram-negative bacilli and examined by Gram staining for both sputum adequacy (using the quality [Q] score) and bacterial morphology. Subjective evaluation of adequacy was also performed and categorized. Based on Q score evaluation, 41% of the samples would have been rejected despite a subjective appearance of purulence. Only three of these rejected samples were culture negative for gram-negative CF pathogens. Correlation between culture results and quantitative Gram stain examination was also poor. These data suggest that subjective evaluation combined with comprehensive bacteriology is superior to Gram staining in identifying pathogens in CF sputum.

Mobile probes

DEFF Research Database (Denmark)

Ørngreen, Rikke; Jørgensen, Anna Neustrup; Noesgaard, Signe Schack

2016-01-01

A project investigating the effectiveness of a collection of online resources for teachers' professional development used mobile probes as a data collection method. Teachers received questions and tasks on their mobile in a dialogic manner while in their everyday context as opposed...... to in an interview. This method provided valuable insight into the contextual use, i.e. how did the online resource transfer to the work practice. However, the research team also found that mobile probes may provide the scaffolding necessary for individual and peer learning at a very local (intra-school) community...... level. This paper is an initial investigation of how the mobile probes process proved to engage teachers in their efforts to improve teaching. It also highlights some of the barriers emerging when applying mobile probes as a scaffold for learning....

Haematoxylin and eosin staining identifies medium to large bacterial aggregates with a reliable specificity: A comparative analysis of follicular bacterial aggregates in axillary biopsies using peptide nucleic acid-fluorescence in situ hybridization and haematoxylin and eosin staining

DEFF Research Database (Denmark)

Ring, Hans Christian; Riis, Peter Theut; Bay, Lene

2017-01-01

between bacterial aggregates identified by haematoxylin and eosin (HE) staining vs bacterial aggregates in corresponding PNA-FISH samples. Axillary biopsies were obtained in 24 healthy controls. HE-stained and PNA-FISH samples were investigated using traditional light microscopy and CLSM, respectively....... The data demonstrate that HE staining identifies large bacterial aggregates (>10 μm) with a sensitivity of 0.43 and specificity of 1. The methods, however, are not equivalent as demonstrated by a McNemar's test (P=.04). Where bacterial aggregates >10 μm in diameter, HE staining may offer a rapid...... and practical low-cost tool to evaluate bacterial aggregates....

Evaluation of surviving fraction using nonclonogenic staining densitometry method

International Nuclear Information System (INIS)

Nishiguchi, Iku; Ogawa, Koichi; Ito, Hisao; Hashimoto, Shozo

1994-01-01

This study was performed to compare our nonclonogenic survival assay (densitometry assay, DM assay) with the widely used clonogenic assay. The established cell lines (HaLa, RMUG, IMR, GOTO) were grown in F 10 medium. The cells were spread in 24-well plates, irradiated with different doses, cultured for about one week and stained with crystal violet after the culture period. Taking the transparent images of the stained well on the light source with the CCD camera, the images were collected with the matrix size 64 x 64, and the integrated optical density of the entire surface of each well was determined by computer with our original program. As the number of cells in the well is reflected by its staining density, the surviving fraction was calculated as the fraction of growth in the irradiated wells relative to controls. The survival curves obtained by the densitometry method showed good correlations with those obtained by clonogenic assay. It is possible to predict intrinsic radiosensitivity with this assay, even if the cells do not form good colonies. However, this method is based on measurements in cultures which depend on the metabolism and growth kinetics of the irradiated cells. Cells should grow exponetially in the same manner in any well to obtain a result similar to that of clonogenic assay, although growth kinetics may be altered by irradiation. This, the endpoint must be strictly standardized. (author)

Alternate gram staining technique using a fluorescent lectin.

Science.gov (United States)

Sizemore, R K; Caldwell, J J; Kendrick, A S

1990-01-01

Fluorescence-labeled wheat germ agglutinin binds specifically to N-acetylglucosamine in the outer peptidoglycan layer of gram-positive bacteria. The peptidoglycan layer of gram-negative bacteria is covered by a membrane and is not labeled by the lectin. By exploiting this phenomenon, an alternative Gram staining technique has been developed. Images PMID:1697149

Accuracy of probing attachment levels using a new computerized cemento-enamel junction probe.

Science.gov (United States)

Deepa, R; Prakash, Shobha

2012-01-01

The assessment of clinical attachment level (CAL) represents the gold standard for diagnosing and monitoring periodontal disease. The aim of the present study was to evaluate the performance of the newly introduced cemento-enamel junction (CEJ) probe in detecting CAL, using CEJ as a fixed reference point, and to compare the CEJ probe with the Florida stent probe (FSP) as well as with a standard manual probe, University of North Carolina-15 (UNC-15). Three examiners recorded the probing attachment level in 384 sites in case group (chronic periodontitis), and in 176 sites, in control group (healthy periodontal status), using the three probes. Subjects included both the sexes and ranged from 35 to 45 years. The experimental design was structured to balance the intra- and inter-examiner consistency at the same site during the two visits. CEJ probe showed higher intra-and inter-examiner consistency over both FSP and UNC-15 in both the case and control groups. Frequency distribution of differences of various magnitudes of repeated measurements ≤1 mm was in the higher range of 86.8% to 87.5% for CEJ probe. The FSP was more reproducible than UNC-15 in detecting relative attachment level (RAL). CEJ automated probe was found to have greatest potential for accuracy and consistency in detecting CAL than FSP and UNC-15. The automated probes appeared to be more reproducible than manual probes.

DNA probes

Energy Technology Data Exchange (ETDEWEB)

Castelino, J

1993-12-31

The creation of DNA probes for detection of specific nucleotide segments differs from ligand detection in that it is a chemical rather than an immunological reaction. Complementary DNA or RNA is used in place of the antibody and is labelled with {sup 32}P. So far, DNA probes have been successfully employed in the diagnosis of inherited disorders, infectious diseases, and for identification of human oncogenes. The latest approach to the diagnosis of communicable and parasitic infections is based on the use of deoxyribonucleic acid (DNA) probes. The genetic information of all cells is encoded by DNA and DNA probe approach to identification of pathogens is unique because the focus of the method is the nucleic acid content of the organism rather than the products that the nucleic acid encodes. Since every properly classified species has some unique nucleotide sequences that distinguish it from every other species, each organism`s genetic composition is in essence a finger print that can be used for its identification. In addition to this specificity, DNA probes offer other advantages in that pathogens may be identified directly in clinical specimens 10 figs, 2 tabs

Complete staining of human spermatozoa and immature germ cells combined with phase contrast microscopy

DEFF Research Database (Denmark)

Michael, A Y; Drejer, J O; Bagger, P V

1987-01-01

A method combining Janus green B and Thymol blue stains the anterior part of the head, the nuclear membrane, middle piece, and tail of spermatozoa light green and the nucleus deep purple. The method provides excellent stained preparations for the evaluation of sperm morphology by phase contrast...

Standardization of fixation, processing and staining methods for the central nervous system of vertebrates.

Science.gov (United States)

Aldana Marcos, H J; Ferrari, C C; Benitez, I; Affanni, J M

1996-12-01

This paper reports the standardization of methods used for processing and embedding various vertebrate brains of different size in paraffin. Other technical details developed for avoiding frequent difficulties arising during laboratory routine are also reported. Some modifications of the Nissl and Klüver-Barrera staining methods are proposed. These modifications include: 1) a Nissl stain solution with a rapid and efficient action with easier differentiation; 2) the use of a cheap microwave oven for the Klüver-Barrera stain. These procedures have the advantage of permitting Nissl and Klüver-Barrera staining of nervous tissue in about five and fifteen minutes respectively. The proposed procedures have been tested in brains obtained from fish, amphibians, reptiles and mammals of different body sizes. They are the result of our long experience in preparing slides for comparative studies. Serial sections of excellent quality were regularly obtained in all the specimens studied. These standardized methods, being simple and quick, are recommended for routine use in neurobiological laboratories.

Microbiology of liver abscesses and the predictive value of abscess gram stain and associated blood cultures.

Science.gov (United States)

Chemaly, Roy F; Hall, Gerri S; Keys, Thomas F; Procop, Gary W

2003-08-01

Although rare, pyogenic liver abscesses are potentially fatal. We evaluated the predictive value of Gram stain of liver abscess aspirates and temporally associated blood cultures. Gram stains detected bacteria in 79% of the liver abscesses tested. The sensitivity and specificity of Gram stain of the liver abscesses were 90% and 100% for Gram-positive cocci (GPC) and 52% and 94% for Gram-negative bacilli (GNB). The sensitivities of the blood cultures for any GPC and GNB present in the liver abscess were 30% and 39%, respectively. Although, Gram stains and blood cultures offer incomplete detection of the microbial contents of pyogenic liver abscesses, both tests should always accompany liver abscess cultures.

Prophylactic Cefazolin in Amnioinfusions Administered for Meconium-Stained Amniotic Fluid

Directory of Open Access Journals (Sweden)

R. K. Edwards

1999-01-01

Full Text Available Objective: To determine if amnioinfusion with an antibiotic solution decreased the rate of clinical chorioamnionitis and puerperal endometritis in patients with meconium-stained amniotic fluid.

A generally applicable sequential alkaline phosphatase immunohistochemical double staining

NARCIS (Netherlands)

van der Loos, Chris M.; Teeling, Peter

2008-01-01

A universal type of sequential double alkaline phosphatase immunohistochemical staining is described that can be used for formalin-fixed, paraffin-embedded and cryostat tissue sections from human and mouse origin. It consists of two alkaline phosphatase detection systems including enzymatic

Detection of Wolbachia endobacteria in Culex quinquefasciatus by Gimenez staining and confirmation by PCR.

Science.gov (United States)

Muniaraj, M; Paramasivan, R; Sunish, I P; Arunachalam, N; Mariappan, T; Jerald Leo, S Victor; Dhananjeyan, K J

2012-12-01

Wolbachia are common intracellular bacteria that are found in arthropods and nematodes. These endosymbionts are transmitted vertically through host eggs and alter host biology in diverse ways, including the induction of reproductive manipulations, such as feminization, parthenogenesis, male killing and sperm-egg incompatibility. Since they can also move horizontally across species boundaries, Wolbachia is gaining importance in recent days as it could be used as a biological control agent to control vector mosquitoes or for paratransgenic approaches. However, the study of Wolbachia requires sophisticated techniques such as PCR and cell culture facilities which cannot be affordable for many laboratories where the diseases transmitted by arthropod vectors are common. Hence, it would be beneficial to develop a simple method to detect the presence of Wolbachia in arthropods. In this study, we described a method of staining Wolbachia endobacteria, present in the reproductive tissues of mosquitoes. The reliability of this method was compared with Gram staining and PCR based detection. The microscopic observation of the Gimenez stained smear prepared from the teased ovary of wild caught and Wolbachia (+) Cx. quinquefasciatus revealed the presence of pink coloured pleomorphic cells of Wolbachia ranging from cocci, comma shaped cells to bacillus and chain forms. The ovaries of Wolbachia (-) cured mosquito did not show any cell. Although Gram's staining is a reliable differential staining for the other bacteria, the bacterial cells in the smears from the ovaries of wild caught mosquitoes did not take the stain properly and the cells were not clearly visible. The PCR amplified product from the pooled remains of wild caught and Wolbachia (+) Cx. quinquefasciatus showed clear banding, whereas, no banding was observed for the negative control (distilled water) and Wolbachia (-) Cx. quinquefasciatus. The Gimenez staining technique applied, could be used to detect the members of

Noninvasive imaging of multiple myeloma using near infrared fluorescent molecular probe

Science.gov (United States)

Hathi, Deep; Zhou, Haiying; Bollerman-Nowlis, Alex; Shokeen, Monica; Akers, Walter J.

2016-03-01

Multiple myeloma is a plasma cell malignancy characterized by monoclonal gammopathy and osteolytic bone lesions. Multiple myeloma is most commonly diagnosed in late disease stages, presenting with pathologic fracture. Early diagnosis and monitoring of disease status may improve quality of life and long-term survival for multiple myeloma patients from what is now a devastating and fatal disease. We have developed a near-infrared targeted fluorescent molecular probe with high affinity to the α4β1 integrin receptor (VLA-4)overexpressed by a majority of multiple myeloma cells as a non-radioactive analog to PET/CT tracer currently being developed for human diagnostics. A near-infrared dye that emits about 700 nm was conjugated to a high affinity peptidomimmetic. Binding affinity and specificity for multiple myeloma cells was investigated in vitro by tissue staining and flow cytometry. After demonstration of sensitivity and specificity, preclinical optical imaging studies were performed to evaluate tumor specificity in murine subcutaneous and metastatic multiple myeloma models. The VLA-4-targeted molecular probe showed high affinity for subcutaneous MM tumor xenografts. Importantly, tumor cells specific accumulation in the bone marrow of metastatic multiple myeloma correlated with GFP signal from transfected cells. Ex vivo flow cytometry of tumor tissue and bone marrow further corroborated in vivo imaging data, demonstrating the specificity of the novel agent and potential for quantitative imaging of multiple myeloma burden in these models.

Analysis of surface stains on modern gold coins

Energy Technology Data Exchange (ETDEWEB)

Corregidor, V., E-mail: vicky.corregidor@itn.pt [Instituto Tecnológico e Nuclear, Instituto Superior Técnico, Universidade Técnica de Lisboa, E.N. 10, 2686-953 Sacavém (Portugal); CFNUL, Av. Prof. Gama Pinto 2, 1649-003 Lisboa (Portugal); Alves, L.C. [Instituto Tecnológico e Nuclear, Instituto Superior Técnico, Universidade Técnica de Lisboa, E.N. 10, 2686-953 Sacavém (Portugal); CFNUL, Av. Prof. Gama Pinto 2, 1649-003 Lisboa (Portugal); Cruz, J. [Instituto Tecnológico e Nuclear, Instituto Superior Técnico, Universidade Técnica de Lisboa, E.N. 10, 2686-953 Sacavém (Portugal); CFNUL, Av. Prof. Gama Pinto 2, 1649-003 Lisboa (Portugal); Dep. Física, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-216 Caparica (Portugal)

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.

Comparison of Histochemical Staining Methods and Correlation with Transient Elastography in Acute Hepatitis.

Science.gov (United States)

Cabibi, Daniela; Calvaruso, Vincenza; Giuffrida, Letizia; Ingrao, Sabrina; Balsamo, Laura; Giannone, Antonino Giulio; Petta, Salvatore; Di Marco, Vito

2015-03-06

To compare Masson's trichrome (MT), Sirius red (SR) and orcein staining in acute hepatitis (AH) and to correlate them with transient elastography (TE), a noninvasive method to assess hepatic fibrosis. We evaluated liver stiffness by TE in a cohort of 34 consecutive patients and assessed MT-, SR- and orcein-stained biopsies using the METAVIR scoring system and digital image analysis (DIA). MT and SR both showed severe fibrosis (stage III-IV, DIA = 12.7%). Orcein showed absent or mild fibrosis (stage 0-II, DIA = 4.4%; p 12.5 kPa, in keeping with SR/MT but not with orcein results. Even though in AH true elastic fibrosis is typically absent or mild, TE shows elevated stiffness values, in keeping with SR/MT evaluations. If not properly evaluated in the clinical context, these results would lead to an overestimation of fibrosis. Orcein is the only staining able to evidence the absence of true elastic fibrosis, which is a typical feature of AH. This is the first study comparing different staining procedures performed on AH biopsies by DIA versus TE. © 2015 S. Karger AG, Basel.

Probe Storage

NARCIS (Netherlands)

Gemelli, Marcellino; Abelmann, Leon; Engelen, Johannes Bernardus Charles; Khatib, M.G.; Koelmans, W.W.; Zaboronski, Olog; Campardo, Giovanni; Tiziani, Federico; Laculo, Massimo

2011-01-01

This chapter gives an overview of probe-based data storage research over the last three decades, encompassing all aspects of a probe recording system. Following the division found in all mechanically addressed storage systems, the different subsystems (media, read/write heads, positioning, data

The incidence of meconium-stained amniotic fluid from 1980 through 1986, by year and gestational age.

Science.gov (United States)

Dysart, M; Graves, B W; Sharp, E S; Cotsonis, G

1991-09-01

The annual incidence of meconium-stained amniotic fluid was analyzed for changes in a total obstetric sample of 45,115 singleton, vertex, liveborn infants over a 7-year study period. The incidence of meconium-stained amniotic fluid for the total obstetric population was calculated for each year of the study period. The sample was then stratified by estimated gestational age, and the incidence of meconium-stained amniotic fluid was calculated for each gestational age group. The incidence of meconium-stained amniotic fluid increased 40.9% over the study period, from 18.8% in 1980 to 26.5% in 1986 (P less than .001). This increase was found to be in a consistent linear trend (P less than .05). The incidence of meconium-stained amniotic fluid was also found to increase significantly in a linear trend as gestational age of the fetus increased. These findings lend support to both the maturational theory and the stress theory of meconium passage in utero.

Simple and rapid staining for detection of Entamoeba cysts and other protozoans with fluorochromes.

Science.gov (United States)

Kawamoto, F; Mizuno, S; Fujioka, H; Kumada, N; Sugiyama, E; Takeuchi, T; Kobayashi, S; Iseki, M; Yamada, M; Matsumoto, Y

1987-02-01

Three fluorochromes were applied to stain various parasitic protozoans. By double staining with 4',6-diamidino-2-phenylindole and propidium iodide, differentiation of the nuclei from the cytoplasm can easily be achieved within several seconds. The chromatoid bodies in Entamoeba cysts were stained bright red. Plasmodium yoelii at all stages except late trophozoites and young gametocytes was easily identified. In the oocysts of Cryptosporidium sp., the nuclei and cytoplasm of the sporozoites fluoresced bluish white and red, respectively, whereas the residual body appeared blue or green. The third fluorochrome, Calcofluor white M2R, was suitable for detecting the cysts of Entamoeba spp. and Chilomastix mesnili.

Interobserver reproducibility and accuracy of p16/Ki-67 dual-stain cytology in cervical cancer screening.

Science.gov (United States)

Wentzensen, Nicolas; Fetterman, Barbara; Tokugawa, Diane; Schiffman, Mark; Castle, Philip E; Wood, Shannon N; Stiemerling, Eric; Poitras, Nancy; Lorey, Thomas; Kinney, Walter

2014-12-01

Dual-stain cytology for p16 and Ki-67 has been proposed as a biomarker in cervical cancer screening. The authors evaluated the reproducibility and accuracy of dual-stain cytology among 10 newly trained evaluators. In total, 480 p16/Ki-67-stained slides from human papillomavirus-positive women were evaluated in masked fashion by 10 evaluators. None of the evaluators had previous experience with p16 or p16/Ki-67 cytology. All participants underwent p16/Ki-67 training and subsequent proficiency testing. Reproducibility of dual-stain cytology was measured using the percentage agreement, individual and aggregate κ values, as well as McNemar statistics. Clinical performance for the detection of cervical intraepithelial neoplasia grade 2 or greater (CIN2+) was evaluated for each individual evaluator and for all evaluators combined compared with the reference evaluation by a cytotechnologist who had extensive experience with dual-stain cytology. The percentage agreement of individual evaluators with the reference evaluation ranged from 83% to 91%, and the κ values ranged from 0.65 to 0.81. The combined κ value was 0.71 for all evaluators and 0.73 for cytotechnologists. The average sensitivity and specificity for the detection of CIN2+ among novice evaluators was 82% and 64%, respectively; whereas the reference evaluation had 84% sensitivity and 63% specificity, respectively. Agreement on dual-stain positivity increased with greater numbers of p16/Ki-67-positive cells on the slides. Good to excellent reproducibility of p16/Ki-67 dual-stain cytology was observed with almost identical clinical performance of novice evaluators compared with reference evaluations. The current findings suggest that p16/Ki-67 dual-stain evaluation can be implemented in routine cytology practice with limited training. © 2014 American Cancer Society.

Persepsi Pemustaka Terhadap Kualitas Layanan Perpustakaan Pascasarjana STAIN Pamekasan

Directory of Open Access Journals (Sweden)

Hairul A Cahyo

2014-07-01

Full Text Available Library is one of supporting element from institute, which can fullfill user’s information need for teaching learning process especially in postgraduate STAIN Pamekasan. To create optimal and good service in postgraduate library, it can see from user’s perception in service quality in postgraduate STAIN Pamekasan. This research used data collection technic; observation, interview and documentation. Officer quality in giving servive to user visible in copability and attitude from officer it self. Library service also can see from collection of books in library to fullfill user’s need. Supporting tools in building and rooms. To complete some tools, it also need infrastructure to fullfill library needs in service. Infrastructure needed are utensils, it means to support library actinity which unused up such as; bookshelf, table and chair, computer and internet, air conditioner, room’s light or lamp, catalogue and photocopy machine.

Removing foxing stains from old paper at 157 nm

International Nuclear Information System (INIS)

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

2003-01-01

Using a molecular fluorine laser at 157 nm foxing stains were removed successfully from a 16th century old paper. Laser cleaning of stains and foxing from old paper manuscripts is far more effective at 157 nm in comparison to different wavelengths without leaving any yellowish after-effect on the paper. This is because at 157 nm illumination of old paper, complete bond breaking of all the organic molecules of the paper is taking place. Mass spectroscopy at 157 nm and for moderate laser intensities up to 1 mJ/cm 2 of old paper suffering from foxing indicate organic matter disintegration to small photofragments atomic, diatomic or triatomic, which are flying apart with supersonic speed. In addition high spatial resolution energy dispersive X-ray system (EDXS) analysis over the effected areas indicate the presence of iron, suggesting that biological activity is taking place preferentially in paper areas containing iron

Intrapartum amnioinfusion in meconium-stained liquor: a case-control study.

Science.gov (United States)

Bansal, Neeta; Gupta, Vineeta; Nanda, Anuja; Chaudhary, Priyanka; Tandon, Archna; Behl, Neelima

2013-06-01

The aim of this study was to investigate perinatal outcome and the rate of cesarean section (CS) following intrapartum amnioinfusion in women with meconium-stained amniotic fluid (MSAF). A total of 100 women at term in labor with meconium were randomized to infuse transcervical intrapartum amnioinfusion with saline (50) and routine obstetrical care (50). Perinatal outcome and obstetric outcome were recorded and analyzed in both groups by means of Chi-square test. The CS rate due to fetal distress was 40.0 % in the control group and 20.0 % in the study group. The difference was statistically significant (P Amnioinfusion in cases of meconium-stained liquor significantly improved neonatal outcome and CS rate without increasing any maternal and fetal complications.

False-Negative Rate of Gram-Stain Microscopy for Diagnosis of Septic Arthritis: Suggestions for Improvement

Directory of Open Access Journals (Sweden)

Paul Stirling

2014-01-01

Full Text Available We quantify the false-negative diagnostic rate of septic arthritis using Gram-stain microscopy of synovial fluid and compare this to values reported in the peer-reviewed literature. We propose a method of improving the diagnostic value of Gram-stain microscopy using Lithium Heparin containers that prevent synovial fluid coagulation. Retrospective study of the Manchester Royal Infirmary microbiology database of patients undergoing synovial fluid Gram-stain and culture between December 2003 and March 2012 was undertaken. The initial cohort of 1896 synovial fluid analyses for suspected septic arthritis was reduced to 143 after exclusion criteria were applied. Analysis of our Gram-stain microscopy yielded 111 false-negative results from a cohort size of 143 positive synovial fluid cultures, giving a false-negative rate of 78%. We report a false-negative rate of Gram-stain microscopy for septic arthritis of 78%. Clinicians should therefore avoid the investigation until a statistically significant data set confirms its efficacy. The investigation's value could be improved by using Lithium Heparin containers to collect homogenous synovial fluid samples. Ongoing research aims to establish how much this could reduce the false-negative rate.

Development of techniques using DNA analysis method for detection/analysis of radiation-induced mutation. Development of an useful probe/primer and improvement of detection efficacy

International Nuclear Information System (INIS)

Maekawa, Hideaki; Tsuchida, Kozo; Hashido, Kazuo; Takada, Naoko; Kameoka, Yosuke; Hirata, Makoto

1999-01-01

Previously, it was demonstrated that detection of centromere became easy and reliable through fluorescent staining by FISH method using a probe of the sequence preserved in α-satelite DNA. Since it was, however, found inappropriate to detect dicentrics based on the relative amount of DNA probe on each chromosome. A prove which allows homogeneous detection of α-satelite DNA for each chromosome was constructed. A presumed sequence specific to kinetochore, CENP-B box was amplified by PCR method and the product DNA was used as a probe. However, the variation in amounts of probe DNA among chromosomes was decreased by only about 20%. Then, a program for image processing of the results obtained from FISH using α-satelite DNA was constructed to use as a marker for centromere. When compared with detection of abnormal chromosomes stained by the conventional method, calculation efficacy for only detection of centromere was improved by the use of this program. Calculation to discriminate the normal or not was still complicated and the detection efficacy was little improved. Chromosomal abnormalities in lymphocytes were used to detect the effects of radiation. In this method, it is needed to shift the phase of cells into metaphase. The mutation induced by radiation might be often repaired during shifting. To exclude this possibility, DNA extraction was conducted at a low temperature and immediately after exposure to 137 Cs, and a rapid genome detection method was established using the genome DNA. As the model genomes, the following three were used: 1) long chain repeated sequences widely dispersed over chromosome, 2) cluster genes, 3) single copy genes. The effects of radiation were detectable at 1-2 Gy for the long repeated sequences and at 7 Gy for the cluster genes, respectively, whereas no significant effects were observed at any Gy tested for the single copy genes. Amplification was marked in the cells exposed at 1-10 Gy (peak at 4 Gy), suggesting that these regions had

Pre-staining thin layer chromatography method for amino acid ...

African Journals Online (AJOL)

Jane

2010-12-13

Dec 13, 2010 ... inexpensive and the results obtained were clean and reproducible. However, it is suitable for the high throughput screening of amino acid-producing strains. Key words: Thin layer chromatography, pre-staining, amino acid detection. INTRODUCTION. Several analytical techniques have been often used for.

Interlaboratory variability of Ki67 staining in breast cancer

NARCIS (Netherlands)

Focke, Cornelia M.; Bürger, Horst; van Diest, Paul J.; Finsterbusch, Kai; Gläser, Doreen; Korsching, Eberhard; Decker, Thomas; Anders, M.; Bollmann, R.; Eiting, Fr; Friedrich, K.; Habeck, J. O.; Haroske, G.; Hinrichs, B.; Behrens, A.; Krause, Lars Udo; Braun-Lang, U.; Lorenzen, J.; Minew, N.; Mlynek-Kersjes, M.; Nenning, H.; Packeisen, J.; Poche-de Vos, F.; Reyher-Klein, S.; Rothacker, D.; Schultz, M.; Sturm, U.; Tawfik, M.; Berghäuser, K. H.; Böcker, W; Cserni, G.; Habedank, S.; Lax, S.; Moinfar, F.; Regitnig, P.; Reiner-Concin, A.; Rüschoff, J.; Varga, Z.; Woziwodski, J.

2017-01-01

Background Postanalytic issues of Ki67 assessment in breast cancers like counting method standardisation and interrater bias have been subject of various studies, but little is known about analytic variability of Ki67 staining between pathology labs. Our aim was to study interlaboratory variability

Sperm viability assessment in marine invertebrates by fluorescent staining and spectrofluorimetry: A promising tool for assessing marine pollution impact.

Science.gov (United States)

Gallo, Alessandra; Boni, Raffaele; Tosti, Elisabetta

2018-01-01

The viability of spermatozoa is a crucial parameter to evaluate their quality that is an important issue in ecotoxicological studies. Here, a new method has been developed to rapidly determine the viability of spermatozoa in three marine invertebrates: the ascidian Ciona intestinalis, the sea urchin Paracentrotus lividus and the mollusc Mytilus galloprovincialis. This method employed the dual DNA fluorescent staining coupled with spectrofluorimetric analysis. The dual fluorescent staining used the SYBR-14 stained live spermatozoa and propidium iodide stained degenerated cells that had lost membrane integrity. Stain uptake was assessed by confocal microscopy and then the percentage of live and dead spermatozoa was quantified by spectrofluorimetric analysis. The microscopic examination revealed three populations of spermatozoa: living-SYBR-14 stained, dead-PI stained, and dying-doubly stained spermatozoa. The fluorescence emission peak values recorded in a spectrofluorimeter provide the portion of live and dead spermatozoa showing a significant negative correlation. The stain combination was further validated using known ratios of live and dead spermatozoa. The present study demonstrated that the dual DNA staining with SYBR-14 and propidium iodide was effective in assessing viability of spermatozoa in marine invertebrates and that spectrofluorimetric analysis can be successfully employed to evaluate the percentage of live and dead spermatozoa. The method develop herein is simple, accurate, rapid, sensitive, and cost-effective, so it could be a useful tool by which marine pollutants may be screened for spermiotoxicity. Copyright © 2017 Elsevier Inc. All rights reserved.

Red-staining of the wall rock and its influence on the reducing capacity around water conducting fractures

International Nuclear Information System (INIS)

Drake, Henrik; Tullborg, Eva-Lena; Annersten, Hans

2008-01-01

Red-staining and alteration of wall rock is common around water conducting fractures in the Laxemar-Simpevarp area (SE Sweden), which is currently being investigated by the Swedish Nuclear Fuel and Waste Management Co. (SKB) in common with many other places. Red-staining is often interpreted as a clear sign of oxidation but relevant analyses are seldom performed. The area is dominated by Palaeoproterozoic crystalline rocks ranging in composition from quartz monzodiorite to granite. In this study wall rock samples have been compared with reference samples from within 0.1 to 1 m of the red-stained rock, in order to describe mineralogical and geochemical changes but also changes in redox conditions. A methodology for tracing changes in mineralogy, mineral and whole rock chemistry and Fe 3+ /Fe tot ratio in silicates and oxides in the red-stained wall rock and the reference rock is reported. The results show that the red-stained rock adjacent to the fractures displays major changes in mineralogy; biotite, plagioclase and magnetite have been altered and chlorite, K-feldspar, albite, sericite, prehnite, epidote and hematite have been formed. The changes in chemistry are however moderate; K-enrichment, Ca-depletion and constant Fe tot are documented. The Fe 3+ /Fe tot ratio in the oxide phase is higher in the red-stained samples whereas the Fe 3+ /Fe tot ratio in the silicate phase is largely similar in the wall rock and the reference samples. Because most of the Fe is hosted in the silicate phase the decrease in reducing capacity (Fe 2+ ), if any, in the red-stained wall rock is very small and not as high as macroscopic observations might suggest. Instead, the mineralogical changes in combination with the modest oxidation and formation of minute hematite grains in porous secondary minerals in pseudomorphs after plagioclase have produced the red-staining. Increased porosity is also characteristic for the red-stained rock. Moderate alteration in the macroscopically fresh

Modular Rake of Pitot Probes

Science.gov (United States)

Dunlap, Timothy A.; Henry, Michael W.; Homyk, Raymond P.

2004-01-01

The figure presents selected views of a modular rake of 17 pitot probes for measuring both transient and steady-state pressures in a supersonic wind tunnel. In addition to pitot tubes visible in the figure, the probe modules contain (1) high-frequency dynamic-pressure transducers connected through wires to remote monitoring circuitry and (2) flow passages that lead to tubes that, in turn, lead to remote steady-state pressure transducers. Prior pitot-probe rakes were fabricated as unitary structures, into which the individual pitot probes were brazed. Repair or replacement of individual probes was difficult, costly, and time-consuming because (1) it was necessary to remove entire rakes in order to unbraze individual malfunctioning probes and (2) the heat of unbrazing a failed probe and of brazing a new probe in place could damage adjacent probes. In contrast, the modules in the present probe are designed to be relatively quickly and easily replaceable with no heating and, in many cases, without need for removal of the entire rake from the wind tunnel. To remove a malfunctioning probe, one first removes a screw-mounted V-cross-section cover that holds the probe and adjacent probes in place. Then one removes a screw-mounted cover plate to gain access to the steady-state pressure tubes and dynamicpressure wires. Next, one disconnects the tube and wires of the affected probe. Finally, one installs a new probe in the reverse of the aforementioned sequence. The wire connections can be made by soldering, but to facilitate removal and installation, they can be made via miniature plugs and sockets. The connections between the probe flow passages and the tubes leading to the remote pressure sensors can be made by use of any of a variety of readily available flexible tubes that can be easily pulled off and slid back on for removal and installation, respectively.

QUANTITATIVE STUDY OF GASTRIC EPITHELIAL LESIONS BY NUCLEOLAR ORGANIZER REGION STAINING

Directory of Open Access Journals (Sweden)

M.R. Arab

2004-11-01

Full Text Available Nucleolar organizer regions (NOR are defined as nucleolar components containing a set of argyrophilic proteins which are selectively stained by colloidal silver nitrate staining. Although studies have shown that the number of NOR dots or particles is directly related to the rapidity of cell proliferation in cancer cells, prognostic or diagnostic value of NOR remains controversial. The aim of the present study was to asses the proliferative activity of the NOR in different gastric epithelial lesions. For these purposes 60 biopsy and surgical specimens of stomach from pathology files of Khatamalanbia and Imam Hospitals were chosen. For each patient, 3-5 paraffin sections were prepared and stained by one step colloidal silver nitrate solution. In each section intranuclear dots in 100 cell nuclei were counted by two of authors in randomly selected fields and data were analyzed by ANOVA. Statistical analysis showed significant difference for NOR number between gastritis, different grades of dysplasia and carcinoma. The shape and number of NOR showed a grater variability in carcinoma compared to other lesions. It seems that NOR could reflect the proliferative activity of cells.

Interkoneksi Stain, Bsm, Dan Mes Dalam Mengembangkan Ekonomi Islam: Studi Kasus Di Purwokerto

OpenAIRE

Dahlan, Ahmad

2008-01-01

STAIN, Bank Syariah Mandiri (BSM), and Masyarakat Ekonomi Syariah (MES, Syariah Economic Society) have important rolein the development of Islam economy in Purwokerto. This research result evidenced that these three institutions have an Islamic economydevelopment program according with vision and mission of their institution. STAIN as Islamic higher education is on curriculum design andlecture quality domain, BSM as banking financial institution on enhancing employee/manager quality, and MES ...

INTERKONEKSI STAIN, BSM, DAN MES DALAM MENGEMBANGKAN EKONOMI ISLAM: STUDI KASUS DI PURWOKERTO

OpenAIRE

Dahlan, Ahmad

2015-01-01

STAIN, Bank Syariah Mandiri (BSM), and Masyarakat Ekonomi Syariah (MES, Syariah Economic Society) have important rolein the development of Islam economy in Purwokerto. This research result evidenced that these three institutions have an Islamic economydevelopment program according with vision and mission of their institution. STAIN as Islamic higher education is on curriculum design andlecture quality domain, BSM as banking financial institution on enhancing employee/manager quality, and MES ...

[Comet assay of DNA fragmentation: modification of silver staining for obtaining permanent preparations].

Science.gov (United States)

Kamins'kyĭ, V O; Lutsyk, M D; Stoĭka, R S

2005-01-01

Modification of comet analysis is proposed for obtaining permanent preparations by DNA staining with silver compounds. The sensitivity of staining is similar to that observed at the treatment by ethidium bromide and other fluorochromes. The advantages of the method are stability of slides and possibility of their reinvestigation by light microscopy. The method does not need expensive fluorescent microscope and lacks contacting with carcinogenic compounds and UV light irradiation.

STM-SQUID probe microscope

International Nuclear Information System (INIS)

Hayashi, Tadayuki; Tachiki, Minoru; Itozaki, Hideo

2007-01-01

We have developed a STM-SQUID probe microscope. A high T C SQUID probe microscope was combined with a scanning tunneling microscope for investigation of samples at room temperature in air. A high permeability probe needle was used as a magnetic flux guide to improve the spatial resolution. The probe with tip radius of less than 100 nm was prepared by microelectropolishing. The probe was also used as a scanning tunneling microscope tip. Topography of the sample surface could be measured by the scanning tunneling microscope with high spatial resolution prior to observation by SQUID microscopy. The SQUID probe microscope image could be observed while keeping the distance from the sample surface to the probe tip constant. We observed a topographic image and a magnetic image of Ni fine pattern and also a magnetically recorded hard disk. Furthermore we have investigated a sample vibration method of the static magnetic field emanating from a sample with the aim of achieving a higher signal-to-noise (S/N) ratio

Characterization of coating probe with Ti-DLC for electrical scanning probe microscope

International Nuclear Information System (INIS)

Shia Xiaolei; Guo Liqiu; Bai Yang; Qiao Lijie

2011-01-01

In electrical scanning probe microscope (ESPM) applications, the wear and conductivity of the probe are undoubtedly serious concerns since they affect the integrity of the measurements. This study investigates the characterization of Ti doped diamond-like-carbon (DLC) as coating material on a silicon cantilever for ESPM. We deposited a layer of Ti-DLC thin film on the surface of Si cantilever by magnetron sputtering. The morphology and composition of the Ti-DLC films were characterized by scanning electron microscopy and Raman spectroscopy, respectively. We also compared the wear resistance, electric conductivity and scanning image quality of the Ti-DLC-coated probes with those of commercially available conductive probes. The results showed that the electric conductivity and the scanning image quality of the Ti-DLC-coated probes were the same as the commercial conductive probes, while the wear resistance and service life was significantly better.

Effects of a Baking Soda Gum on extrinsic dental stain: results of a longitudinal 4-week assessment.

Science.gov (United States)

Soparkar, P; Newman, M B

2001-07-01

An evaluation of the effects of ARM & HAMMER DENTAL CARE The Baking Soda Gum (AHDC) on extrinsic dental stain was made in 48 subjects presenting with measurable extrinsic stain. The subjects were randomized to use either the baking soda gum or a non-baking soda placebo gum for 20 minutes twice daily after lunch and dinner while brushing once daily. The procedure of limited brushing was chosen to simulate the level of hygiene normally practiced by participants entering a clinical study. After 4 weeks, the reduction in measurable extrinsic stain in the baking soda gum group was statistically significant (P = .0044) relative to baseline. Statistical analysis of the placebo gum group revealed no significant change in extrinsic stain from baseline. The magnitude of the unadjusted longitudinal reduction in extrinsic stain in the baking soda gum group was 29.7% at 4 weeks.

[Immunocytochemical demonstration of astrocytes in brain sections combined with Nissl staining].

Science.gov (United States)

Korzhevskiĭ, D E; Otellin, V A

2004-01-01

The aim of the present study was to develop an easy and reliable protocol of combined preparation staining, which would unite the advantages of immunocytochemical demonstration of astrocytes with the availability to evaluate functional state of neurons provided by Nissl technique. The presented protocol of paraffin sections processing allows to retain high quality of tissue structure and provides for selective demonstration of astrocytes using the monoclonal antibodies against glial fibrillary acidic protein and contrast Nissl staining of cells. The protocol can be used without any changes for processing of brain sections obtained from the humans and other mammals with the exception of mice and rabbits.

Taste in Art-Exposure to Histological Stains Shapes Abstract Art Preferences.

Science.gov (United States)

Böthig, Antonia M; Hayn-Leichsenring, Gregor U

2017-01-01

Exposure to art increases the appreciation of artworks. Here, we showed that this effect is domain independent. After viewing images of histological stains in a lecture, ratings increased for restricted subsets of abstract art images. In contrast, a lecture on art history generally enhanced ratings for all art images presented, while a lecture on town history without any visual stimuli did not increase the ratings. Therefore, we found a domain-independent exposure effect of images of histological stains to particular abstract paintings. This finding suggests that the 'taste' for abstract art is altered by visual impressions that are presented outside of an artistic context.

Measuring the Level of Agreement Between Cloacal Gram's Stains and Bacterial Cultures in Hispaniolan Amazon Parrots ( Amazona ventralis ).

Science.gov (United States)

Evans, Erika E; Mitchell, Mark A; Whittington, Julia K; Roy, Alma; Tully, Thomas N

2014-12-01

Cloacal or fecal Gram's stains and bacterial cultures are routinely performed during avian physical examinations to assess the microbial flora of the gastrointestinal tract. Although cloacal or fecal Gram's stains and bacterial cultures are considered routine diagnostic procedures, the level of agreement between the individual tests has not been determined. To investigate the level of agreement between results from Gram's stain and bacterial culture when used to assess cloacal or fecal samples from psittacine birds, samples were taken from 21 clinically healthy Hispaniolan Amazon parrots ( Amazona ventralis ) and tested by Gram's stain cytology and bacterial culture. Most bacteria (97.2%) identified by Gram's stain were gram positive. However, gram-negative organisms were identified in 7 of 21 (33.3%; 95% confidence interval: 13.3%-53.3%) birds. Escherichia coli was the only gram-negative organism identified on culture. Agreement between results of Gram's stain and culture was fair (weighted κ = 0.27). The results of this study suggest that Gram's stains and bacterial culture may need to be performed with a parallel testing strategy to limit the likelihood of misclassifying the microbial flora of psittacine patients.

[Standardization of Blastocystis hominis diagnosis using different staining techniques].

Science.gov (United States)

Eymael, Dayane; Schuh, Graziela Maria; Tavares, Rejane Giacomelli

2010-01-01

The present study was carried out from March to May 2008, with the aim of evaluating the effectiveness of different techniques for diagnosing Blastocystis hominis in a sample of the population attended at the Biomedicine Laboratory of Feevale University, Novo Hamburgo, Rio Grande do Sul. On hundred feces samples from children and adults were evaluated. After collection, the samples were subjected to the techniques of spontaneous sedimentation (HPJ), sedimentation in formalin-ether (Ritchie) and staining by means of Gram and May-Grünwald-Giemsa (MGG). The presence of Blastocystis hominis was observed in 40 samples, when staining techniques were used (MGG and Gram), while sedimentation techniques were less efficient (32 positive samples using the Ritchie technique and 20 positive samples using the HPJ technique). Our results demonstrate that HPJ was less efficient than the other methods, thus indicating the need to include laboratory techniques that enable parasite identification on a routine basis.

Proximal Probes Facility

Data.gov (United States)

Federal Laboratory Consortium — The Proximal Probes Facility consists of laboratories for microscopy, spectroscopy, and probing of nanostructured materials and their functional properties. At the...

Accelerated identification of proteins by mass spectrometry by employing covalent pre-gel staining with Uniblue A.

Directory of Open Access Journals (Sweden)

Marco A Mata-Gómez

Full Text Available BACKGROUND: The identification of proteins by mass spectrometry is a standard method in biopharmaceutical quality control and biochemical research. Prior to identification by mass spectrometry, proteins are usually pre-separated by electrophoresis. However, current protein staining and de-staining protocols are tedious and time consuming, and therefore prolong the sample preparation time for mass spectrometry. METHODOLOGY AND PRINCIPAL FINDINGS: We developed a 1-minute covalent pre-gel staining protocol for proteins, which does not require de-staining before the mass spectrometry analysis. We investigated the electrophoretic properties of derivatized proteins and peptides and studied their behavior in mass spectrometry. Further, we elucidated the preferred reaction of proteins with Uniblue A and demonstrate the integration of the peptide derivatization into typical informatics tools. CONCLUSIONS AND SIGNIFICANCE: The Uniblue A staining method drastically speeds up the sample preparation for the mass spectrometry based identification of proteins. The application of this chemo-proteomic strategy will be advantageous for routine quality control of proteins and for time-critical tasks in protein analysis.

Validation of a Fully Automated HER2 Staining Kit in Breast Cancer

Directory of Open Access Journals (Sweden)

Cathy B. Moelans

2010-01-01

Full Text Available Background: Testing for HER2 amplification and/or overexpression is currently routine practice to guide Herceptin therapy in invasive breast cancer. At present, HER2 status is most commonly assessed by immunohistochemistry (IHC. Standardization of HER2 IHC assays is of utmost clinical and economical importance. At present, HER2 IHC is most commonly performed with the HercepTest which contains a polyclonal antibody and applies a manual staining procedure. Analytical variability in HER2 IHC testing could be diminished by a fully automatic staining system with a monoclonal antibody.

[1,10]Phenanthroline based cyanine dyes as fluorescent probes for ribonucleic acids in live cells

Science.gov (United States)

Kovalska, Vladyslava; Kuperman, Marina; Varzatskii, Oleg; Kryvorotenko, Dmytro; Kinski, Elisa; Schikora, Margot; Janko, Christina; Alexiou, Christoph; Yarmoluk, Sergiy; Mokhir, Andriy

2017-12-01

A series of monomethine, trimethine- and styrylcyanine dyes based on a [1,10]phenanthroline moiety was synthesized, characterized and investigated as potential fluorescent probes for nucleic acids in cell free settings and in cells. The dyes were found to be weakly fluorescent in the unbound state, whereas upon the binding to dsDNA or RNA their emission intensity raised up to 50 times (for monomethine benzothiazole derivative FT1 complexed with RNA). The strongest fluorescence intensity in assemblies with dsDNA and RNA was observed for the trimethine benzothiazole derivative FT4. The quantum yield of FT4 fluorescence in its complex with dsDNA was found to be 1.5% and the binding constant (K b) was estimated to be 7.9 × 104 M-1 that is a typical value for intercalating molecules. The FT4 dye was found to be cell membrane permeable. It stains RNA rich components—the nucleoli and most probably the cytoplasmic RNA. FT4 bound to RNAs delivers a very strong fluorescence signal, which makes this easily accessible dye a potentially useful alternative to known RNA stains, e.g. expensive SYTO® 83. The advantage of FT4 is its easy synthetic access including no chromatographic purification steps, which will be reflected in its substantially lower price.

Double immunohistochemical staining with laminin 5 (γ2 chain) and collagen IV in colorectal neoplasms

DEFF Research Database (Denmark)

Fiehn, Anne-Marie Kanstrup; Bzorek, Michael; Warnecke, Mads

2016-01-01

divided according to the primary histopathological diagnoses of tubular adenoma, tubulovillous adenoma, adenoma with pseudoinvasion and glandular adenocarcinoma stages pT1, pT2 or pT3, were included in the study. In normal colonic mucosa, no expression of laminin 5 staining was observed. BM was always...... as a supplement for the diagnosis of pT1 CRC. In adenomas, the double staining highlights the areas for the pathologist to pay extra attention. By itself, the double staining cannot determine whether or not there is invasion. Morphology remains the single most important factor in differentiating adenoma...

‘Resurrecting Harry Clarke’: Breathing life into stained glass tourism in Ireland

Directory of Open Access Journals (Sweden)

Tony Kiely

2014-12-01

Full Text Available Internationally, the exponential demand for ‘cultural/heritage’ tourism is increasingly being viewed by tourism stakeholders as an opportunity for value adding revenue generation, wherein both specialist and ‘media programmed’ tourists can seek out designated cultural attractions to satisfy their respective quests for authentic, and/or emotionally charged experiences. Indeed, this international ‘demand’ re-alignment is exemplified in the growth of churches and cathedrals who openly promote their artistic content as ‘must see attractions’. However, despite such utilitarian attractiveness, one wonders if the counter-influences of indifference, protectionism, or fear of heritage commodification, might act to scupper an opportunity to re-envision Harry Clarke’s iconic stained glass church windows as tourist attractions in Ireland? Born in Dublin in 1889, into a city consumed with the heady mix of Catholic Emancipation and artistic rebirth, Harry Clarke grew to pre-eminence among international stained glass artists at the turn of the twentieth century. Yet, despite being universally acclaimed as a master of stained glass creativity, his oeuvre, from a tourism perspective, has been sadly overlooked, with most promotion of Ireland’s twentieth century cultural renaissance being focused both on its literary giants (Joyce, Wilde, Swift, Beckett, Yeats and Stoker, and its traditional music culture. However, with the attendant economic spinoff offered by religious and cultural tourism growth, one wonders if the ubiquitous presence in Irish churches of Harry Clarke’s ecclesiastical stained glass windows might be re-imagined by core and peripheral stakeholders as marketable tourist attractions? Employing a qualitative methodology, involving semi structured interviews with key informants, this paper explores stakeholder awareness of Clarke’s significance as a potential tourist attractor, in addition to the appropriateness of promoting

Enamels in stained glass windows: Preparation, chemical composition, microstructure and causes of deterioration

International Nuclear Information System (INIS)

Schalm, O.; Van der Linden, V.; Frederickx, P.; Luyten, S.; Van der Snickt, G.; Caen, J.; Schryvers, D.; Janssens, K.; Cornelis, E.; Van Dyck, D.; Schreiner, M.

2009-01-01

Stained glass windows incorporating dark blue and purple enamel paint layers are in some cases subject to severe degradation while others from the same period survived the ravages of time. A series of dark blue, green-blue and purple enamel glass paints from the same region (Northwestern Europe) and from the same period (16-early 20th centuries) has been studied by means of a combination of microscopic X-ray fluorescence analysis, electron probe micro analysis and transmission electron microscopy with the aim of better understanding the causes of the degradation. The chemical composition of the enamels diverges from the average chemical composition of window glass. Some of the compositions appear to be unstable, for example those with a high concentration of K 2 O and a low content of CaO and PbO. In other cases, the deterioration of the paint layers was caused by the less than optimal vitrification of the enamel during the firing process. Recipes and chemical compositions indicate that glassmakers of the 16-17th century had full control over the color of the enamel glass paints they made. They mainly used three types of coloring agents, based on Co (dark blue), Mn (purple) and Cu (light-blue or green-blue) as coloring elements. Blue-purple enamel paints were obtained by mixing two different coloring agents. The coloring agent for red-purple enamel, introduced during the 19th century, was colloidal gold embedded in grains of lead glass.

Comparison of various staining techniques in the diagnosis of ...

African Journals Online (AJOL)

Journal of Medical and Biomedical Sciences ... This can be achieved via various diagnostic techniques, commonly microscopy in this environment, hence the need to compare the efficacy of the commonly ... The objective of the study is to identify the most effective of the commonly used stains in identifying these parasites.

Development of conductivity probe and temperature probe for in-situ measurements in hydrological studies

International Nuclear Information System (INIS)

Chandra, U.; Galindo, B.J.; Castagnet, A.C.G.

1981-05-01

A conductivity probe and a temperature probe have been developed for in-situ measurements in various hydrological field studies. The conductivity probe has platinum electrodes and is powered with two 12 volt batteries. The sensing element of the temperature probe consists of a resistor of high coefficient of temperature. Response of the conductivity probe is measured in a milliampere mater while the resistance of the thermistor is read by a digital meter. The values of conductivity and temperature are derived from respective calibration. The probes are prototype and their range of measurement can be improved depending upon the requirement of the field problem. (Author) [pt

Laser-heated emissive plasma probe.

Science.gov (United States)

Schrittwieser, Roman; Ionita, Codrina; Balan, Petru; Gstrein, Ramona; Grulke, Olaf; Windisch, Thomas; Brandt, Christian; Klinger, Thomas; Madani, Ramin; Amarandei, George; Sarma, Arun K

2008-08-01

Emissive probes are standard tools in laboratory plasmas for the direct determination of the plasma potential. Usually they consist of a loop of refractory wire heated by an electric current until sufficient electron emission. Recently emissive probes were used also for measuring the radial fluctuation-induced particle flux and other essential parameters of edge turbulence in magnetized toroidal hot plasmas [R. Schrittwieser et al., Plasma Phys. Controlled Fusion 50, 055004 (2008)]. We have developed and investigated various types of emissive probes, which were heated by a focused infrared laser beam. Such a probe has several advantages: higher probe temperature without evaporation or melting and thus higher emissivity and longer lifetime, no deformation of the probe in a magnetic field, no potential drop along the probe wire, and faster time response. The probes are heated by an infrared diode laser with 808 nm wavelength and an output power up to 50 W. One probe was mounted together with the lens system on a radially movable probe shaft, and radial profiles of the plasma potential and of its oscillations were measured in a linear helicon discharge.

Laser-heated emissive plasma probe

International Nuclear Information System (INIS)

Schrittwieser, Roman; Ionita, Codrina; Balan, Petru; Gstrein, Ramona; Grulke, Olaf; Windisch, Thomas; Brandt, Christian; Klinger, Thomas; Madani, Ramin; Amarandei, George; Sarma, Arun K.

2008-01-01

Emissive probes are standard tools in laboratory plasmas for the direct determination of the plasma potential. Usually they consist of a loop of refractory wire heated by an electric current until sufficient electron emission. Recently emissive probes were used also for measuring the radial fluctuation-induced particle flux and other essential parameters of edge turbulence in magnetized toroidal hot plasmas [R. Schrittwieser et al., Plasma Phys. Controlled Fusion 50, 055004 (2008)]. We have developed and investigated various types of emissive probes, which were heated by a focused infrared laser beam. Such a probe has several advantages: higher probe temperature without evaporation or melting and thus higher emissivity and longer lifetime, no deformation of the probe in a magnetic field, no potential drop along the probe wire, and faster time response. The probes are heated by an infrared diode laser with 808 nm wavelength and an output power up to 50 W. One probe was mounted together with the lens system on a radially movable probe shaft, and radial profiles of the plasma potential and of its oscillations were measured in a linear helicon discharge

Laser-heated emissive plasma probe

Science.gov (United States)

Schrittwieser, Roman; Ionita, Codrina; Balan, Petru; Gstrein, Ramona; Grulke, Olaf; Windisch, Thomas; Brandt, Christian; Klinger, Thomas; Madani, Ramin; Amarandei, George; Sarma, Arun K.

2008-08-01

Emissive probes are standard tools in laboratory plasmas for the direct determination of the plasma potential. Usually they consist of a loop of refractory wire heated by an electric current until sufficient electron emission. Recently emissive probes were used also for measuring the radial fluctuation-induced particle flux and other essential parameters of edge turbulence in magnetized toroidal hot plasmas [R. Schrittwieser et al., Plasma Phys. Controlled Fusion 50, 055004 (2008)]. We have developed and investigated various types of emissive probes, which were heated by a focused infrared laser beam. Such a probe has several advantages: higher probe temperature without evaporation or melting and thus higher emissivity and longer lifetime, no deformation of the probe in a magnetic field, no potential drop along the probe wire, and faster time response. The probes are heated by an infrared diode laser with 808nm wavelength and an output power up to 50W. One probe was mounted together with the lens system on a radially movable probe shaft, and radial profiles of the plasma potential and of its oscillations were measured in a linear helicon discharge.

Role of intrapartum transcervical amnioinfusion in patients with meconium-stained amniotic fluid.

Science.gov (United States)

Bhatia, Pushpa; Reena, Kumari; Nangia, Sangita

2013-03-01

The study was undertaken to evaluate maternal, perinatal outcomes following transcervical intrapartum amnioinfusion in women with meconium-stained amniotic fluid. A prospective comparative study was conducted on 100 women with meconium-stained amniotic fluid in labor. Group A: study group (50 cases) received amnioinfusion. Group B: control group (50 cases) did not receive amnioinfusion. FHR monitoring was done using cardiotocography. Significant relief from variable decelerations was seen in 68.18 % cases in the amnioinfusion group as compared to 7.1 % cases in the control group. 78 % cases who were given amnioinfusion had vaginal delivery as compared to 18 % cases in the control group. Fourteen percent cases in the study group had cesarean delivery as compared to 68 % cases in the control group. Meconium aspiration syndrome was seen in six percent neonates in the study group as compared to 20 % in the control group. Two neonates died in the control group due to meconium aspiration syndrome. There was no maternal mortality or major maternal complication. Intrapartum transcervical amnioinfusion is valuable in patients with meconium-stained amniotic fluid.

Colour stabilities of three types of orthodontic clear aligners exposed to staining agents

OpenAIRE

Liu, Chen-Lu; Sun, Wen-Tian; Liao, Wen; Lu, Wen-Xin; Li, Qi-Wen; Jeong, Yunho; Liu, Jun; Zhao, Zhi-He

2016-01-01

The aim of this study was to evaluate and compare the colour stabilities of three types of orthodontic clear aligners exposed to staining agents in vitro. Sixty clear orthodontic aligners produced by three manufacturers (Invisalign, Angelalign, and Smartee) were immersed in three staining solutions (coffee, black tea, and red wine) and one control solution (distilled water). After 12-h and 7-day immersions, the aligners were washed in an ultrasonic cleaner and measured with a colourimeter. Th...

Fluorescent pH-Sensing Probe Based on Biorefinery Wood Lignosulfonate and Its Application in Human Cancer Cell Bioimaging.

Science.gov (United States)

Xue, Yuyuan; Liang, Wanshan; Li, Yuan; Wu, Ying; Peng, Xinwen; Qiu, Xueqing; Liu, Jinbin; Sun, Runcang

2016-12-28

A water-soluble, ratiometric fluorescent pH probe, L-SRhB, was synthesized via grafting spirolactam Rhodamine B (SRhB) to lignosulfonate (LS). As the ring-opening product of L-SRhB, FL-SRhB was also prepared. The pH-response experiment indicated that L-SRhB showed a rapid response to pH changes from 4.60 to 6.20 with a pK a of 5.35, which indicated that L-SRhB has the potential for pH detection of acidic organelle. In addition, the two probes were internalized successfully by living cells through the endocytosis pathway and could distinguish normal cells from cancer cells by different cell staining rates. In addition, L-SRhB showed obvious cytotoxicity to cancer cells, whereas it was nontoxic to normal cells in the same condition. L-SRhB might have potential in cancer therapy. L-SRhB might be a promising ratiometric fluorescent pH sensor and bioimaging dye for the recognition of cancer cells. The results also provided a new perspective to the high-value utilization of lignin.

Double side multicrystalline silicon passivation by one step stain etching-based porous silicon

Energy Technology Data Exchange (ETDEWEB)

Mohamed, Seifeddine Belhadj; Ben Rabha, Mohamed; Bessais, Brahim [Laboratoire de Photovoltaique, Centre de Recherches et des Technologies de l' Energie, Technopole de Borj-Cedria, BP 95, 2050 Hammam-Lif (Tunisia)

2012-10-15

In this paper, we investigate the effect of stain etching-based porous silicon on the double side multicrystalline silicon. Special attention is given to the use of the stain etched PS as an antireflection coating as well as for surface passivating capabilities. Stain etching of double side multicrystalline silicon leads to the formation of PS nanostructures, that dramatically decrease the surface reflectivity from 30% to about 7% and increase the effective lifetime from 1 {mu}s to 10 {mu}s at a minority carrier density ({Delta}n) of 10{sup 15} cm{sup -3}. These results let us correlate the rise of the lifetime values to the photoluminescence intensity to the hydrogen and oxide passivation as shown by FTIR analysis. This low-cost PS formation process can be applied in the photovoltaic cell technology as a standard procedure (copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

X-ray induction of micronuclei in human lymphocyte subpopulations differentiated by immunoperoxidase staining

International Nuclear Information System (INIS)

Ban, Sadayuki; Nakano, Mimako; Cologne, J.B.

1992-10-01

In this report we sought to confirm the radiosensitivity of human peripheral blood lymphocyte subpopulations using a micronucleus assay. Mononucleated cells isolated from peripheral blood were irradiated with X rays. After being cultured for 3 days, cells were fixed and stained using the immunoperoxidase staining technique. Lymphocyte subpopulations were characterized by means of the monoclonal antibodies Leu4 (CD3), Leu2a (CD8), and Leu19 (CD56). Dose-response curves were obtained by scoring the number of micronuclei in binucleated cells that reacted with a specific antibody and were then stained. The dose response of CD8 + (suppressor/cytotoxic) cells was quite similar to that of CD3 + (pan T) cells. In comparison, CD56 + (natural killer) cells were significantly less sensitive, although scorable binucleated CD56 + cells made up less than 4 % of the total number of binucleated cells. (author)

Hemangiomas versus arterio-venous malformations: Role of elastic stains and mast cell density

Directory of Open Access Journals (Sweden)

Pravin Pawane

2014-01-01

Full Text Available Context: Vascular anomalies present diagnostic challenges to histopathologists. Mulliken and Glowacki′s classification categorizes vascular lesions into hemangiomas and vascular malformations. Aim: This study explored diagnostic clues in the histomorphology of hemangiomas and vascular malformations. Materials and Methods: A total of 120 cases of benign vascular lesions were retrieved from 12 years period. A total of 94 cases, where complete clinical details and representative paraffin sections were available, were included in this study. Hematoxylin and eosin (H and E stain and Verhoeff′s stain for elastic tissue were done on all cases and lesions were classified into hemangiomas or arterio-venous malformations (AVM. Mast cell density in all lesions was calculated from toluidine blue stained sections. Results: Ten cases of hemangiomas were reclassified as AVM on the basis of presence and absence of arteriovenous structures. Intra-lesional nerves were seen in significantly higher number of AVMs compared to hemangiomas. Medium and thick sized nerve bundles were seen in 56% of AVMs, while they were not seen in any of the hemangiomas. Mean mast cell density was significantly higher in proliferating hemangiomas (53.12 ± 27.83 cells/mm 2 compared to involuting hemangiomas (11.43 ± 7.9 cells/mm 2 . Conclusions: Use of elastic tissue stains are useful ancillary tools to distinguish between AVMs and hemangiomas. The presence of arteries and arterioles are an integral part of AVMs. The presence of the intra-lesional nerves can be useful to distinguish between AVMs and hemangiomas even on H and E stained sections. The significantly higher mast cell density seen in proliferating hemangiomas compared with involuting ones, seem to suggest that mast cells play an important role in the natural history of these lesions.

Innovative SPM Probes for Energy-Storage Science: MWCNT-Nanopipettes to Nanobattery Probes

Science.gov (United States)

Larson, Jonathan; Talin, Alec; Pearse, Alexander; Kozen, Alexander; Reutt-Robey, Janice

As energy-storage materials and designs continue to advance, new tools are needed to direct and explore ion insertion/de-insertion at well-defined battery materials interfaces. Scanned probe tips, assembled from actual energy-storage materials, permit SPM measures of local cathode-anode (tip-sample) interactions, including ion transfer. We present examples of ``cathode'' MWCNT-terminated STM probe tips interacting with Li(s)/Si(111) anode substrates. The MWCNT tip functions as both SPM probe and Li-nanopipette,[1] for controlled transport and manipulation of Li. Local field conditions for lithium ionization and transfer are determined and compared to electrostatic models. Additional lithium metallic and oxide tips have been prepared by thin film deposition on conventional W tips, the latter of which effectively functions as a nanobattery. We demonstrate use of these novel probe materials in the local lithiation of low-index Si anode interfaces, probing local barriers for lithium insertion. Prospects and limitations of these novel SPM probes will be discussed. U.S. Department of Energy Award Number DESC0001160.

Characterisation of the nucleolar organising regions during the cell cycle in two varieties of Petunia hybrida as visualised by fluorescence in situ hybridisation and silver staining.

Science.gov (United States)

Montijn, M B; ten Hoopen, R; Fransz, P F; Oud, J L; Nanninga, N

1998-05-01

The cell cycle-dependent spatial position, morphology and activity of the four nucleolar organising regions (NORs) of the Petunia hybrida cultivar Mitchell and the inbred line V26 have been analysed. Application of the silver staining technique and fluorescence in situ hybridisation on fixed root-tip material revealed that these interspecific hybrids possess four NORs of which only those of chromosome 2 are active during interphase, which implies that the NOR activity is not of parental origin. However, at the end of mitosis, activity of all NOR regions could be detected, suggesting that the high demand for ribosomes at this stage of the cell cycle requires temporal activity of all NORs. Using actin DNA probes as markers in fluorescence in situ hybridisation experiments enabled the identification of the individual petunia chromosomes.

Developmental venous anomalies with capillary stain: a subgroup of symptomatic DVAs?

International Nuclear Information System (INIS)

Roccatagliata, Luca; Berg, Rene van den; Soderman, Michael; Boulin, Anne; Condette-Auliac, Stephanie; Rodesch, Georges

2012-01-01

Intracranial developmental venous anomalies (DVAs) are considered benign vascular dispositions; they are asymptomatic in the vast majority of cases. They represent extreme variations of the venous drainage and may rarely be responsible for focal venous ischemia leading to neurological dysfunction. The aim of the study is to analyze a group of patients with symptomatic DVAs with capillary stain at angiography. We retrospectively reviewed the clinical and radiological features of patients in which a DVA was considered the cause of a neurological event. In all the patients, the DVA was suspected by angio-CT or MRI and conventional angiography was performed to detail the angioarchitecture of the DVA. A total of 7 patients and 11 DVAs were identified; three patients had multiple DVAs. Three DVAs were frontal, two were parietal, two were thalamic, one was in the midbrain, and three were cerebellar. Patients presented with progressive neurological deficits, seizures, or cerebral hemorrhage. All these DVAs were associated with a peculiar capillary stain at angiography. Although being normal anatomical variations, DVAs may create, because of hemodynamic unbalance, venous ischemia that induces angiogenic phenomena. MRI shows the suffering of the brain and angiography witnesses this angiogenesis under the form of capillary stain. Conventional angiography can thus provide useful information to recognize ''atypical'' symptomatic DVAs. (orig.)

Intrapartum amnioinfusion for meconium-stained fluid: meta-analysis of prospective clinical trials.

Science.gov (United States)

Pierce, J; Gaudier, F L; Sanchez-Ramos, L

2000-06-01

To evaluate the effectiveness of intrapartum prophylactic amnioinfusion in pregnancies complicated by meconium-stained amniotic fluid. We identified prospective clinical trials of amnioinfusion in pregnancies complicated by meconium-stained amniotic fluid (AF) published in English by using computerized databases, references in published studies, and index reviews. We analyzed prospective studies of intrapartum amnioinfusion for meconium-stained AF. In every case, group allocation was based exclusively on meconium in AF. Only published studies with clearly documented outcome data were included. The trials were evaluated for meconium below the vocal cords, meconium aspiration syndrome, fetal acidemia, cesarean delivery, and postpartum endometritis. Each trial was evaluated for the quality of its methodology, inclusion and exclusion criteria, adequacy of randomization, amnioinfusion protocols, definition of outcomes, and statistical analyses. Thirteen studies met inclusion criteria for our systematic review. Odds ratios (ORs) with their 95% confidence intervals (CIs) were calculated for each outcome. Estimates of ORs and risk differences for dichotomous outcomes were calculated using random and fixed-effects models. We tested for homogeneity across the studies. We found that intrapartum amnioinfusion significantly reduced the frequency of meconium aspiration syndrome (OR 0.30; 95% CI 0.19, 0. 46), of meconium below the vocal cords, and neonatal acidemia. Subjects allocated to receive amnioinfusion also had a significantly lower overall cesarean rate (OR 0.74, 95% CI 0.59, 0.93) without increased postpartum endometritis. Amnioinfusion in cases of meconium-stained fluid significantly improves neonatal outcome, lowers the cesarean delivery rate, and does not increase the postpartum endometritis rate.

Kematangan Karir Mahasiswa Jurusan Tarbiyah Sekolah Tinggi Agama Islam Negeri (STAIN Samarinda

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Lina Revilla Malik

2015-06-01

Full Text Available The objective of this study was to describe the level of career readiness of the education faculty students of STAIN Samarinda and the aspects of career readiness of the education faculty students of STAIN Samarinda. The sample of this study was 92 students of the education faculty students. The researcher used stratified random sampling in this study. The data were collected by using questionnaires. The data were analized through rating scale. The result of findings shows that the level of career readiness of the education faculty students of STAIN Samarinda was 73% with 17.156 interval or it is categorized as “Good”. It is affected by the ages of the respondents in the age of 18 – 25 years old which means they are in the sub level of transition and trial little commitment. It is also affected by the factors of interest, skill, and personality. The aspects of career readiness of the education faculty students of STAIN Samarinda are as follows: 1 Career planning is 73.12%, categorized as “Good”. It is affected by the respondents’ knowledge of workfield, interest, education, extra-curricular activities, and spare-times activities. 2 Career exploration is 68.7%, categorized as “Good”. It is affected by the respondents’ family background and social life interaction. 3 Career decision is 76.3%, categorized as “Good”. It is affected by the respondents’ social economical background. 4 Work-field information is 73.5%, categorized as “Good”. It is affected by the educational and economical background of the respondents’ parents.

Color stability of CAD/CAM Zirconia ceramics following exposure to acidic and staining drinks.

Science.gov (United States)

Colombo, Marco; Cavallo, Marco; Miegge, Matteo; Dagna, Alberto; Beltrami, Riccardo; Chiesa, Marco; Poggio, Claudio

2017-11-01

The aim of this in vitro study was to evaluate the color stability of CAD/CAM Zirconia ceramics following exposure to acidic drink (Coca Cola) and after exposure to staining solution (coffee). All the samples were immersed in different staining solutions over a 28-day test period. A colorimetric evaluation according to the CIE L*a*b* system was performed by a blind trained operator at 7, 14, 21, 28 days of the staining process. Shapiro Wilk test and Kruskal-Wallis ANOVA were applied to assess significant differences among restorative materials. Paired t-test was applied to test which CIE L*a*b* parameters significantly changed after immersion in staining solutions. One week immersion in acidic drink did not cause a perceivable discoloration for all restorative materials (ΔE < 3.3). Subsequent immersion in coffee affected color stability of all Zirconia samples, even if Kruskal-Wallis ANOVA found significant differences among the various restorative materials. The ∆Es of CAD/CAM Zirconia ceramics after immersion in coffee varied among the products, but color integrity is not affected by contact with acidic drinks. Key words: CAD/CAM restorative materials, CIE Lab, Zirconia ceramics.

Fluorescence In Situ Hybridization with Peptide Nucleic Acid Probes for Rapid Identification of Candida albicans Directly from Blood Culture Bottles

Science.gov (United States)

Rigby, Susan; Procop, Gary W.; Haase, Gerhard; Wilson, Deborah; Hall, Geraldine; Kurtzman, Cletus; Oliveira, Kenneth; Von Oy, Sabina; Hyldig-Nielsen, Jens J.; Coull, James; Stender, Henrik

2002-01-01

A new fluorescence in situ hybridization (FISH) method that uses peptide nucleic acid (PNA) probes for identification of Candida albicans directly from positive-blood-culture bottles in which yeast was observed by Gram staining (herein referred to as yeast-positive blood culture bottles) is described. The test (the C. albicans PNA FISH method) is based on a fluorescein-labeled PNA probe that targets C. albicans 26S rRNA. The PNA probe is added to smears made directly from the contents of the blood culture bottle and hybridized for 90 min at 55°C. Unhybridized PNA probe is removed by washing of the mixture (30 min), and the smears are examined by fluorescence microscopy. The specificity of the method was confirmed with 23 reference strains representing phylogenetically related yeast species and 148 clinical isolates covering the clinically most significant yeast species, including C. albicans (n = 72), C. dubliniensis (n = 58), C. glabrata (n = 5), C. krusei (n = 2), C. parapsilosis (n = 4), and C. tropicalis (n = 3). The performance of the C. albicans PNA FISH method as a diagnostic test was evaluated with 33 routine and 25 simulated yeast-positive blood culture bottles and showed 100% sensitivity and 100% specificity. It is concluded that this 2.5-h method for the definitive identification of C. albicans directly from yeast-positive blood culture bottles provides important information for optimal antifungal therapy and patient management. PMID:12037084

Theory of NMR probe design

International Nuclear Information System (INIS)

Schnall, M.D.

1988-01-01

The NMR probe is the intrinsic part of the NMR system which allows transmission of a stimulus to a sample and the reception of a resulting signal from a sample. NMR probes are used in both imaging and spectroscopy. Optimal probe design is important to the production of adequate signal/moise. It is important for anyone using NMR techniques to understand how NMR probes work and how to optimize probe design

Probe for intracellular concentrations of drugs: delayed fluorescence from acridine orange

International Nuclear Information System (INIS)

Wardman, P.; Dennis, M.F.; White, J.

1989-01-01

The aim of this work is to develop fluorescent probes that will indicate effective concentrations of therapeutic agents, or endogenous protectors, at important cellular sites. Acridine orange associates with nucleic acids and emits a 'delayed' fluorescence signal. This signal is quenched by oxidants such as oxygen, nitroaryl radiosensitizers, adriamycin and mitomycin-c, and reductants such as thiols, ascorbate and other radioprotectors. The quenching of the acridine orange delayed fluorescence reflects the effective concentration of these therapeutically-important oxidants and reductants near DNA. The relative concentration of basic radiosensitizers such as pimonidazole (Ro 03-8799) near the DNA is greater than that of misonidazole. Thiols quench the delayed fluorescence signal according to the degree of ionization of the thiol function; this may model the reactivity of thiols with guanine radical sites in DNA. Ascorbate and aminopyrine do not quench the delayed fluorescence from cells stained with acridine orange as these compounds are taken up by cells very inefficiently

Christendom's Narratives and the Stained Glass Designs of Yusuf ...

African Journals Online (AJOL)

This paper attempts a recast of Christendom's narratives in the stained glass designs of Yusuf Cameron Adebayo Grillo as the distinctive overarching mechanism of the evangelisation paradigm of the post Vatican II Church. It, therefore, draws attention to the delimitation of time frames in the history of the art form. Using the ...

Evaluation of methods for stain removal in acrylic resin denture teeth: in vitro study

Directory of Open Access Journals (Sweden)

Ana Flávia Balestrero CASSIANO

Full Text Available Abstract Introduction The staining of artificial teeth can be related to the acrylic resin abrasion caused by brushing, resulting in higher deposition of dyes from the beverage, and consequently higher aesthetic damage. Objective The aim of this in vitro study was to evaluate methods for removal of stains from acrylic denture teeth using spectrophotometric analysis. Material and method Artificial teeth were divided into twelve groups (n=10 according to the type of treatment (re-polishing - Re or immersion in Corega Tabs - Sp, staining solutions, coffee (Cf and Coca-Cola® (Cc or water (W and with/without toothbrushing (B. The Sp specimens were submitted to seven immersion cycles (5 min each. The Re specimens were polished with pumice stone followed by Spain white paste. Color differences (ΔE were captured by a spectrophotometer: T0 (baseline, T1 (after brushing/immersion in solutions and T2 (after Re or Sp. Result Statistically significant color change between T1 and T2 (paired T-test; α =.05 was observed for the group CfSp (p=.032; and for the groups BWRe (p=.000, BCfRe (p=.049 and CcRe (p=.042. Higher color changes were observed for the specimens submitted to toothbrushing (ANOVA two way; p<.001. Conclusion It could be concluded that the immersion in sodium perborate (Corega Tabs can be used for removal of coffee stains from denture teeth, and re-polishing for removal of Coca-Cola® stains. Still, toothbrushing produced greater color changes on denture teeth, regardless of the immersion solution.

A nanobody targeting carcinoembryonic antigen as a promising molecular probe for non-small cell lung cancer.

Science.gov (United States)

Wang, Hao; Meng, Ai-Min; Li, Sheng-Hua; Zhou, Xiao-Liang

2017-07-01

Carcinoembryonic antigen (CEA) is a biomarker and therapy target for non‑small cell lung cancer (NSCLC), which is the most common type of lung cancer. Nanobodies with high target specificity are promising candidates to function as anti‑CEA probes. In the present study, the targeting effects of an anti‑CEA nanobody obtained from phage display were investigated using technetium‑99 m (99mTc) and fluorescence labeling. In vitro binding and immunofluorescent staining assays, as well as in vivo blood clearance and biodistribution assays were performed. High specificity and affinity of the nanobody for CEA‑positive H460 cells was observed in vitro. The pharmacokinetics assay of the 99mTc‑nanobody in Wistar rats demonstrated that the nanobody had appropriate T1/2α and T1/2β, which were 20.2 and 143.5 min, respectively. The biodistribution assay using H460 xenograft‑bearing nude mice demonstrated a high ratio of signal in tumor compared with background, which confirmed that the nanobody may be useful as a molecular probe for CEA‑positive cancer, particularly in NSCLC.

Impact of round-the-clock CSF Gram stain on empirical therapy for suspected central nervous system infections.

Science.gov (United States)

Tissot, F; Prod'hom, G; Manuel, O; Greub, G

2015-09-01

The impact of round-the-clock cerebrospinal fluid (CSF) Gram stain on overnight empirical therapy for suspected central nervous system (CNS) infections was investigated. All consecutive overnight CSF Gram stains between 2006 and 2011 were included. The impact of a positive or a negative test on empirical therapy was evaluated and compared to other clinical and biological indications based on institutional guidelines. Bacterial CNS infection was documented in 51/241 suspected cases. Overnight CSF Gram stain was positive in 24/51. Upon validation, there were two false-positive and one false-negative results. The sensitivity and specificity were 41 and 99 %, respectively. All patients but one had other indications for empirical therapy than Gram stain alone. Upon obtaining the Gram result, empirical therapy was modified in 7/24, including the addition of an appropriate agent (1), addition of unnecessary agents (3) and simplification of unnecessary combination therapy (3/11). Among 74 cases with a negative CSF Gram stain and without formal indication for empirical therapy, antibiotics were withheld in only 29. Round-the-clock CSF Gram stain had a low impact on overnight empirical therapy for suspected CNS infections and was associated with several misinterpretation errors. Clinicians showed little confidence in CSF direct examination for simplifying or withholding therapy before definite microbiological results.

Comparison of Histochemical Stainings in Evaluation of Liver Fibrosis and Correlation with Transient Elastography in Chronic Hepatitis

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Daniela Cabibi

2015-01-01

Full Text Available Background and Aim. The best staining to evaluate liver fibrosis in liver hepatitis is still a debated topic. This study aimed to compare Masson’s trichrome (MT, Sirius Red (SR, and orcein stainings in evaluating liver fibrosis in chronic HCV hepatitis (CHC with semiquantitative and quantitative methods (Collagen Proportionate Area (CPA by Digital Image Analysis (DIA and correlate them with transient elastography (TE. Methods. Liver stiffness evaluation of 111 consecutive patients with CHC was performed by TE. Semiquantitative staging by Metavir score system and CPA by DIA were assessed on liver biopsy stained with MT, SR, and orcein. Results. MT, SR, and orcein staining showed concordant results in 89.6% of cases in staging CHC, without significant difference in both semiquantitative and quantitative evaluations of fibrosis. TE values were concordant with orcein levels in 86.5% of the cases and with MT/RS in 77.5% (P<0.001. No significant correlation between the grade of necroinflammatory activity and TE values was found. Conclusion. In CHC, SR/MT and orcein stainings are almost concordant and when discordant, orcein staining is better related to TE values than MT/RS. This suggests that elastic fibers play a more important role than reticular or collagenous ones in determining stiffness values in CHC.

Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain.

Science.gov (United States)

Hezel, Marcus; Ebrahimi, Fahim; Koch, Marco; Dehghani, Faramarz

2012-10-01

Immunohistochemical visualization of antigens in specimen has evolved to an indispensable technique in biomedical research for investigations of cell morphology and pathology both in bright field and fluorescence microscopy. While there are couple of staining methods that reveal entire cytoarchitecture in bright field microscopy such as Nissl or hemalaun-eosin, there are still limitations in visualizations of cytoarchitecture in fluorescence microscopy. The present study reports a simple staining method that provides the required illustration of cell allocations and cellular composition in fluorescence microscopy in adult and in developing rodent central nervous system using the fluorophore propidium iodide (PI, 5μg/mL). PI is a well-accepted marker for degenerating cells when applied prior to fixation (pre-fixation PI staining). Here, PI was added to the sections after the fixation (post-fixation PI staining). This revised labeling procedure led to similar cytoarchitectural staining patterns in fluorescence microscopy as observed with hemalaun in bright field microscopy. This finding was proven in organotypic hippocampal slice cultures (OHSC) and brain sections obtained from different postnatal developmental stages. Excitotoxically lesioned OHSC subjected to pre-fixation PI staining merely showed brightly labeled condensed nuclei of degenerating neurons. In contrast, post-fixation PI staining additionally revealed extensive labeling of neuronal cell bodies and glial cells within the OHSC, thus allowing visualization of stratification of neuronal layers and cell morphology. Furthermore, post-fixation PI staining was combined with NeuN, calbindin, calretinin, glial fibrillary acidic protein or Griffonia simplicifolia isolectin B4 (IB(4)) in post natal (p1 and p9) and adult rats. In early post-natal brain sections almost all mentioned cellular markers led to an incomplete staining of the native cell organization and resulted in an inaccurate estimation of cell

The use of special stains in liver biopsy interpretation: Implications ...

African Journals Online (AJOL)

2015-12-03

Dec 3, 2015 ... Key words: Biliary disease, iron overload, liver biopsy, special stains. Date of Acceptance: 03-Dec- .... effect of cutting fresh sections after trimming. This limits ... alcohol as a significant factor in the iron deposition found in these ...

A comparison study of histochemical staining of various tissues after ...

African Journals Online (AJOL)

mean =5.28) then Carnoy's (mean = 4.00). For Alcian blue and Perl's Prussian blue, the best staining qualities were obtained by Formalin (mean = 4.76 and 5.64 respectively) followed by Carnoy's (mean = 2.88 and 3.92 respectively).

Cultural probes

DEFF Research Database (Denmark)

Madsen, Jacob Østergaard

The aim of this study was thus to explore cultural probes (Gaver, Boucher et al. 2004), as a possible methodical approach, supporting knowledge production on situated and contextual aspects of occupation.......The aim of this study was thus to explore cultural probes (Gaver, Boucher et al. 2004), as a possible methodical approach, supporting knowledge production on situated and contextual aspects of occupation....

Light microscopy with differential staining techniques for the characterisation and discrimination of insects versus marine arthropods processed animal proteins.

Science.gov (United States)

Ottoboni, Matteo; Tretola, Marco; Cheli, Federica; Marchis, Daniela; Veys, Pascal; Baeten, Vincent; Pinotti, Luciano

2017-08-01

The aim of this study was to evaluate the use of light microscopy with differential staining techniques for the discrimination of insect material from marine arthropods - classified as fishmeal. Specifically, three samples of single-species insect material, Hermetia illucens (HI), Bombyx mori (BM) and Tenebrio molitor (TM), and two samples of marine arthropods, shrimp material and krill, were analysed and compared after staining by two reagents to enhance fragment identification. Alizarin Red (AR) and Chlorazol Black (CB), which react respectively with calcium salts and chitin, were tested for their potential efficacy in distinguishing between insect and marine materials. Results indicated that AR failed to stain HI, BM and TM materials. By contrast, the three insect species materials tested were stained by CB. When shrimp fragments and krill were considered, AR and CB stained marine materials reddish-pink and light blue to black, respectively. By combining these results, it can be suggested that CB staining may efficiently be used to mark insect materials; AR does stain shrimp fragments but does not stain the tested insect material, indicating a possible approach for discriminating between insects and marine arthropods. However, since the present study was performed on pure materials and a small set of samples, possible implementation of this technique still needs to be confirmed in complex matrices such as compound feed.

Hepatic steatosis inhibits autophagic proteolysis via impairment of autophagosomal acidification and cathepsin expression

International Nuclear Information System (INIS)

Inami, Yoshihiro; Yamashina, Shunhei; Izumi, Kousuke; Ueno, Takashi; Tanida, Isei; Ikejima, Kenichi; Watanabe, Sumio

2011-01-01

Highlights: → Acidification of autophagosome was blunted in steatotic hepatocytes. → Hepatic steatosis did not disturb fusion of isolated autophagosome and lysosome. → Proteinase activity of cathepsin B and L in autolysosomes was inhibited by steatosis. → Hepatic expression of cathepsin B and L was suppressed by steatosis. -- Abstract: Autophagy, one of protein degradation system, contributes to maintain cellular homeostasis and cell defense. Recently, some evidences indicated that autophagy and lipid metabolism are interrelated. Here, we demonstrate that hepatic steatosis impairs autophagic proteolysis. Though accumulation of autophagosome is observed in hepatocytes from ob/ob mice, expression of p62 was augmented in liver from ob/ob mice more than control mice. Moreover, degradation of the long-lived protein leucine was significantly suppressed in hepatocytes isolated from ob/ob mice. More than 80% of autophagosomes were stained by LysoTracker Red (LTR) in hepatocytes from control mice; however, rate of LTR-stained autophagosomes in hepatocytes were suppressed in ob/ob mice. On the other hand, clearance of autolysosomes loaded with LTR was blunted in hepatocytes from ob/ob mice. Although fusion of isolated autophagosome and lysosome was not disturbed, proteinase activity of cathepsin B and L in autolysosomes and cathepsin B and L expression of liver were suppressed in ob/ob mice. These results indicate that lipid accumulation blunts autophagic proteolysis via impairment of autophagosomal acidification and cathepsin expression.

Hepatic steatosis inhibits autophagic proteolysis via impairment of autophagosomal acidification and cathepsin expression

Energy Technology Data Exchange (ETDEWEB)

Inami, Yoshihiro [Department of Gastroenterology, Juntendo University School of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421 (Japan); Yamashina, Shunhei, E-mail: syamashi@juntendo.ac.jp [Department of Gastroenterology, Juntendo University School of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421 (Japan); Izumi, Kousuke [Department of Gastroenterology, Juntendo University School of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421 (Japan); Ueno, Takashi [Department of Biochemistry, Juntendo University School of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421 (Japan); Tanida, Isei [Department of Biochemistry and Cell Biology, Laboratory of Biomembranes, National Institute of Infectious Disease, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640 (Japan); Ikejima, Kenichi; Watanabe, Sumio [Department of Gastroenterology, Juntendo University School of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421 (Japan)

2011-09-09

Highlights: {yields} Acidification of autophagosome was blunted in steatotic hepatocytes. {yields} Hepatic steatosis did not disturb fusion of isolated autophagosome and lysosome. {yields} Proteinase activity of cathepsin B and L in autolysosomes was inhibited by steatosis. {yields} Hepatic expression of cathepsin B and L was suppressed by steatosis. -- Abstract: Autophagy, one of protein degradation system, contributes to maintain cellular homeostasis and cell defense. Recently, some evidences indicated that autophagy and lipid metabolism are interrelated. Here, we demonstrate that hepatic steatosis impairs autophagic proteolysis. Though accumulation of autophagosome is observed in hepatocytes from ob/ob mice, expression of p62 was augmented in liver from ob/ob mice more than control mice. Moreover, degradation of the long-lived protein leucine was significantly suppressed in hepatocytes isolated from ob/ob mice. More than 80% of autophagosomes were stained by LysoTracker Red (LTR) in hepatocytes from control mice; however, rate of LTR-stained autophagosomes in hepatocytes were suppressed in ob/ob mice. On the other hand, clearance of autolysosomes loaded with LTR was blunted in hepatocytes from ob/ob mice. Although fusion of isolated autophagosome and lysosome was not disturbed, proteinase activity of cathepsin B and L in autolysosomes and cathepsin B and L expression of liver were suppressed in ob/ob mice. These results indicate that lipid accumulation blunts autophagic proteolysis via impairment of autophagosomal acidification and cathepsin expression.

Comparison of modified Chicago sky blue stain and potassium hydroxide mount for the diagnosis of dermatomycoses and onychomycoses.

Science.gov (United States)

Liu, Zhong; Sheng, Ping; Yang, Yan-Ping; Li, Wen; Huang, Wen-Ming; Wang, Jie-Di; Fan, Yi-Ming

2015-05-01

The diagnostic value of modified Chicago sky blue (CSB) stain and potassium hydroxide (KOH) mount for superficial mycoses was compared using fungal culture as gold standard. The sensitivity and screening time of the CSB stain were superior to the KOH mount. The CBS stain is simple, quick and reliable for diagnosing superficial mycoses. Copyright © 2015. Published by Elsevier B.V.

Darkfield illumination improves microscopic detection of metals in Timm's stained tissue

DEFF Research Database (Denmark)

Baatrup, E; Frederickson, C J

1989-01-01

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 undet...... are undetectable in brightfield light microscopy but are easily detected in darkfield microscopy. Darkfield illumination is therefore recommended for improving the detection of trace or toxic metals in tissue. Udgivelsesdato: 1989-Aug......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...

Optical properties of amyloid stained by Congo red: history and mechanisms.

Science.gov (United States)

Howie, Alexander J; Brewer, Douglas B

2009-04-01

Amyloid stained by Congo red has striking optical properties that generally have been poorly described and inadequately explained, although they can be understood from principles of physical optics. Molecules of Congo red are orientated on amyloid fibrils, and so the dye becomes dichroic and birefringent. The birefringence varies with wavelength in accordance with a fundamental property of all light-transmitting materials called anomalous dispersion of the refractive index around an absorption peak. The combination of this and absorption of light, with modification by any additional birefringence in the optical system, explains the various colours that can be seen in Congo red-stained amyloid between crossed polariser and analyser, and also when the polariser and analyser are progressively uncrossed. These are called anomalous colours.

Dual-probe decoherence microscopy: probing pockets of coherence in a decohering environment

International Nuclear Information System (INIS)

Jeske, Jan; Cole, Jared H; Müller, Clemens; Marthaler, Michael; Schön, Gerd

2012-01-01

We study the use of a pair of qubits as a decoherence probe of a nontrivial environment. This dual-probe configuration is modelled by three two-level systems (TLSs), which are coupled in a chain in which the middle system represents an environmental TLS. This TLS resides within the environment of the qubits and therefore its coupling to perturbing fluctuations (i.e. its decoherence) is assumed much stronger than the decoherence acting on the probe qubits. We study the evolution of such a tripartite system including the appearance of a decoherence-free state (dark state) and non-Markovian behaviour. We find that all parameters of this TLS can be obtained from measurements of one of the probe qubits. Furthermore, we show the advantages of two qubits in probing environments and the new dynamics imposed by a TLS that couples to two qubits at once. (paper)

Evaluation of gram-chromotrope kinyoun staining technique: its effectiveness in detecting microsporidial spores in fecal specimens.

Science.gov (United States)

Salleh, Fatmah M; Al-Mekhlafi, Abdulsalam M; Nordin, Anisah; Yasin, 'Azlin M; Al-Mekhlafi, Hesham M; Moktar, Norhayati

2011-01-01

This study was conducted to evaluate the modification of the usual Gram-chromotrope staining technique developed in-house known as Gram-chromotrope Kinyoun (GCK) in comparison with the Weber Modified Trichrome (WMT) staining technique; as the reference technique. Two hundred and ninety fecal specimens received by the Microbiology Diagnostic Laboratory of Hospital Universiti Kebangsaan Malaysia were examined for the presence of microsporidial spores. The sensitivity and specificity of GCK compared to the reference technique were 98% and 98.3%, respectively. The positive and negative predictive values were 92.5% and 99.6%, respectively. The agreement between the reference technique and the GCK staining technique was statistically significant by Kappa statistics (K = 0.941, P staining technique has high sensitivity and specificity in the detection of microsporidial spores in fecal specimens. Hence, it is recommended to be used in the diagnosis of intestinal microsporidiosis. Copyright © 2011 Elsevier Inc. All rights reserved.

Phase-selective staining of metal salt for scanning electron microscopy imaging of block copolymer film

Energy Technology Data Exchange (ETDEWEB)

Li, Jing Ze, E-mail: Lijinge@uestc.edu.cn [State Key Laboratory of Electronic Thin Films and Integrated Devices, School of Microelectronic and Solid-state Electronic, University of Electronic Science and Technology of China, Chengdu 610054 (China); State Key Laboratory of Polymer Materials Engineering (Sichuan University), Chengdu 610054 (China); Xinjiang Key Laboratory of Electronic Information Materials and Devices, Urumuqi 830011 (China); Wang, Ying; Hong Wang, Zhi; Mei, Di; Zou, Wei [State Key Laboratory of Electronic Thin Films and Integrated Devices, School of Microelectronic and Solid-state Electronic, University of Electronic Science and Technology of China, Chengdu 610054 (China); Min Chang, Ai [State Key Laboratory of Polymer Materials Engineering (Sichuan University), Chengdu 610054 (China); Wang, Qi [Xinjiang Key Laboratory of Electronic Information Materials and Devices, Urumuqi 830011 (China); Komura, Motonori; Ito, Kaori [Division of Integrated Molecular Engineering, Chemical Resources Laboratory, Tokyo Institute of Technology, Yokohama 226-8503 (Japan); Iyoda, Tomokazu, E-mail: Iyoda.t.aa@m.titech.ac.jp [Division of Integrated Molecular Engineering, Chemical Resources Laboratory, Tokyo Institute of Technology, Yokohama 226-8503 (Japan)

2010-09-15

Three metal salts, i.e., AgNO{sub 3}, HAuCl{sub 4}, and KCl, were proposed as novel staining reagents instead of traditional RuO{sub 4} and OsO{sub 4} labeled with expensive price and extreme toxicity for scanning electron microscopy (SEM) imaging of microphase separated block copolymer film. A simple and costless aqueous solution immersion procedure could ensure selective staining of the metal slat in specific phase of the nanostructured copolymer film, leading to a clear phase contrasted SEM image. The heavy metal salt has better staining effect, demonstrating stable and high signal-to-noise SEM image even at an acceleration voltage as high as 30 kV and magnification up to 250,000 times.

Resolution enhancement of pump-probe microscope with an inverse-annular filter

Science.gov (United States)

Kobayashi, Takayoshi; Kawasumi, Koshi; Miyazaki, Jun; Nakata, Kazuaki

2018-04-01

Optical pump-probe microscopy can provide images by detecting changes in probe light intensity induced by stimulated emission, photoinduced absorbance change, or photothermal-induced refractive index change in either transmission or reflection mode. Photothermal microscopy, which is one type of optical pump-probe microscopy, has intrinsically super resolution capability due to the bilinear dependence of signal intensity of pump and probe. We introduce new techniques for further resolution enhancement and fast imaging in photothermal microscope. First, we introduce a new pupil filter, an inverse-annular pupil filter in a pump-probe photothermal microscope, which provides resolution enhancement in three dimensions. The resolutions are proved to be improved in lateral and axial directions by imaging experiment using 20-nm gold nanoparticles. The improvement in X (perpendicular to the common pump and probe polarization direction), Y (parallel to the polarization direction), and Z (axial direction) are by 15 ± 6, 8 ± 8, and 21 ± 2% from the resolution without a pupil filter. The resolution enhancement is even better than the calculation using vector field, which predicts the corresponding enhancement of 11, 8, and 6%. The discussion is made to explain the unexpected results. We also demonstrate the photothermal imaging of thick biological samples (cells from rabbit intestine and kidney) stained with hematoxylin and eosin dye with the inverse-annular filter. Second, a fast, high-sensitivity photothermal microscope is developed by implementing a spatially segmented balanced detection scheme into a laser scanning microscope using a Galvano mirror. We confirm a 4.9 times improvement in signal-to-noise ratio in the spatially segmented balanced detection compared with that of conventional detection. The system demonstrates simultaneous bi-modal photothermal and confocal fluorescence imaging of transgenic mouse brain tissue with a pixel dwell time of 20 µs. The

A case of perforated sigmoid diverticulitis in which gram staining of ascitic fluid was useful for diagnosis.

Science.gov (United States)

Tsuchida, Junko; Fujita, Shouhei; Kawano, Fumihiro; Tsukamoto, Ryoichi; Honjo, Kunpei; Naito, Shigetoshi; Ishiyama, Shun; Miyano, Shozo; Machida, Michio; Kitabatake, Toshiaki; Fujisawa, Minoru; Kojima, Kuniaki; Ogura, Kanako; Matsumoto, Toshiharu

2014-01-01

An 85-year-old woman was admitted to our hospital for steroid therapy for relapsing nephrotic syndrome. During hospitalization, she complained of sudden epigastric pain at night. Although there were signs of peritoneal irritation, CT showed a large amount of ascitic fluid, but no free intraperitoneal gas. Gram staining of ascitic fluid obtained by abdominal paracentesis showed Gram-negative rods, which raised a strong suspicion of gastrointestinal perforation and peritonitis. Therefore, emergency surgery was performed. Exploration of the colon showed multiple sigmoid diverticula, one of which was perforated. The patient underwent an emergency Hartmann's procedure. Imaging studies failed to reveal any evidence of gastrointestinal perforation, presenting a diagnostic challenge. However, a physician performed rapid Gram staining of ascitic fluid at night when laboratory technicians were absent, had a strong suspicion of gastrointestinal perforation, and performed emergency surgery. Gram staining is superior in rapidity, and ascitic fluid Gram staining can aid in diagnosis, suggesting that it should be actively performed. We report this case, with a review of the literature on the significance of rapid diagnosis by Gram staining.

Urgensi Redesain Kebijakan dan Sistem Penyelenggaraan Studi Keislaman pada Peskam STAIN Samarinda

Directory of Open Access Journals (Sweden)

M. Said Husin

2014-06-01

Full Text Available This study explores the urgent of STAIN Samarinda Boarding College Program (PESKAM to redesign both its policy and its instructional framework for the irregular Islamic studies system, academic year 2014/2015, viewed from various aspects. The first aspect is about the effectiveness of irregular Islamic studies instructional achievement run in (academic year 2103/2014. The second aspect is the leaders of STAIN Samarinda political interests concerning with PESKAM empowerment including institutional, instructional design, financial suffort, and the staffs. The third aspect is the regular Islamic studies lecturers’ academic sensitivity. And the last aspect is concerning with the design reliability of irregular Islamic studies program. The research shows that redesign is a must. It is imperative for STAIN Samarinda to redesign both PESKAM policy and its instructional framework for the irregular Islamic studies system. First, the system run during academic year 2013/2014 could not be considered effective. It did not use any general academic standards and could not be approached in terms of modern instructional management and design, such as the evaluation system designed to know periodically both the competence that a learner (a student in PESKAM must be about to master and the process run to do such instruction as well.  However, all staffs and those who were in charge of managing the program worked hard and did all their best. Second, irregular Islamic studies instruction is viewed as the spirit for PESKAM in general. Its valuable benefit meets the stakeholders’ need. Finally, it is the cornerstone which can be accredited as the prime aspect distinguishing STAIN Samarinda from any general university (PTN/PTS

Model for resonant plasma probe.

Energy Technology Data Exchange (ETDEWEB)

Warne, Larry Kevin; Johnson, William Arthur; Hebner, Gregory Albert; Jorgenson, Roy E.; Coats, Rebecca Sue

2007-04-01

This report constructs simple circuit models for a hairpin shaped resonant plasma probe. Effects of the plasma sheath region surrounding the wires making up the probe are determined. Electromagnetic simulations of the probe are compared to the circuit model results. The perturbing effects of the disc cavity in which the probe operates are also found.

High-definition hematoxylin and eosin staining in a transition to digital pathology

Directory of Open Access Journals (Sweden)

Jamie D Martina

2011-01-01

Full Text Available Introduction: A lot of attention has been generated in recent years by digital pathology and telepathology. Multiple reasons for and barriers to effective adoption are discussed in the current literature. Digital slides are the most promising medium at this time. The goal of our study was to evaluate whether the change in the methodology, particularly utilizing the so-called high-definition hematoxylin and eosin (H and E slides, enhanced the quality of the final digital slide, and whether pathologists who tested the results perceived this as a difference in quality. Methods: The study was a blinded comparison of digital slides prepared using two methods: standard H&E batch staining and automated individual "high definition" HD HE staining. Four pathologists have compared 80 cases stained with each method. Results: The results discussed in this study show potential promise that the utilization of protocol(s adapted for tissue and for imaging might be preferable for digital pathology in at least some of the pathology subspecialties. In particular, the protocol evaluated here was capable of turning out digital slides that had more contrast and detail, and therefore were perceived to provide enhanced diagnostically significant information for the pathologist.

Color stability of CAD/CAM Zirconia ceramics following exposure to acidic and staining drinks

Science.gov (United States)

Colombo, Marco; Cavallo, Marco; Miegge, Matteo; Dagna, Alberto; Beltrami, Riccardo; Chiesa, Marco

2017-01-01

Background The aim of this in vitro study was to evaluate the color stability of CAD/CAM Zirconia ceramics following exposure to acidic drink (Coca Cola) and after exposure to staining solution (coffee). Material and Methods All the samples were immersed in different staining solutions over a 28-day test period. A colorimetric evaluation according to the CIE L*a*b* system was performed by a blind trained operator at 7, 14, 21, 28 days of the staining process. Shapiro Wilk test and Kruskal-Wallis ANOVA were applied to assess significant differences among restorative materials. Paired t-test was applied to test which CIE L*a*b* parameters significantly changed after immersion in staining solutions. Results One week immersion in acidic drink did not cause a perceivable discoloration for all restorative materials (ΔE < 3.3). Subsequent immersion in coffee affected color stability of all Zirconia samples, even if Kruskal-Wallis ANOVA found significant differences among the various restorative materials. Conclusions The ∆Es of CAD/CAM Zirconia ceramics after immersion in coffee varied among the products, but color integrity is not affected by contact with acidic drinks. Key words:CAD/CAM restorative materials, CIE Lab, Zirconia ceramics. PMID:29302281

A rapid method combining Golgi and Nissl staining to study neuronal morphology and cytoarchitecture.

Science.gov (United States)

Pilati, Nadia; Barker, Matthew; Panteleimonitis, Sofoklis; Donga, Revers; Hamann, Martine

2008-06-01

The Golgi silver impregnation technique gives detailed information on neuronal morphology of the few neurons it labels, whereas the majority remain unstained. In contrast, the Nissl staining technique allows for consistent labeling of the whole neuronal population but gives very limited information on neuronal morphology. Most studies characterizing neuronal cell types in the context of their distribution within the tissue slice tend to use the Golgi silver impregnation technique for neuronal morphology followed by deimpregnation as a prerequisite for showing that neuron's histological location by subsequent Nissl staining. Here, we describe a rapid method combining Golgi silver impregnation with cresyl violet staining that provides a useful and simple approach to combining cellular morphology with cytoarchitecture without the need for deimpregnating the tissue. Our method allowed us to identify neurons of the facial nucleus and the supratrigeminal nucleus, as well as assessing cellular distribution within layers of the dorsal cochlear nucleus. With this method, we also have been able to directly compare morphological characteristics of neuronal somata at the dorsal cochlear nucleus when labeled with cresyl violet with those obtained with the Golgi method, and we found that cresyl violet-labeled cell bodies appear smaller at high cellular densities. Our observation suggests that cresyl violet staining is inadequate to quantify differences in soma sizes.

Patterns of Gram-stained fecal flora as a quick diagnostic marker in patients with severe SIRS.

Science.gov (United States)

Shimizu, Kentaro; Ogura, Hiroshi; Tomono, Kazunori; Tasaki, Osamu; Asahara, Takashi; Nomoto, Koji; Morotomi, Masami; Matsushima, Asako; Nakahori, Yasutaka; Yamano, Shuhei; Osuka, Akinori; Kuwagata, Yasuyuki; Sugimoto, Hisashi

2011-06-01

The gut is an important target organ of injury during critically ill conditions. Although Gram staining is a common and quick method for identifying bacteria, its clinical application has not been fully evaluated in critically ill conditions. This study's aims were to identify patterns of Gram-stained fecal flora and compare them to cultured bacterial counts and to investigate the association between the patterns and septic complications in patients with severe systemic inflammatory response syndrome (SIRS). Fifty-two patients with SIRS were included whose Gram-stained fecal flora was classified into three patterns. In a diverse pattern, large numbers of multiple kinds of bacteria completely covered the field. In a single pattern, one specific kind of bacteria or fungi predominantly covered the field. In a depleted pattern, most bacteria were diminished in the field. In the analysis of fecal flora, the numbers of total obligate anaerobes in the depleted pattern was significantly lower than those in the diverse pattern and single pattern (p Gram-stained fecal flora can be classified into three patterns and are associated with both cultured bacterial counts and clinical information. Gram-stained fecal bacteria can be used as a quick bedside diagnostic marker for severe SIRS patients.

Characterization of near-field optical probes

DEFF Research Database (Denmark)

Vohnsen, Brian; Bozhevolnyi, Sergey I.

1999-01-01

Radiation and collection characteristics of four different near-field optical-fiber probes, namely, three uncoated probes and an aluminium-coated small-aperture probe, are investigated and compared. Their radiation properties are characterized by observation of light-induced topography changes...... in a photo-sensitive film illuminated with the probes, and it is confirmed that the radiated optical field is unambigiously confined only for the coated probe. Near-field optical imaging of a standing evanescent-wave pattern is used to compare the detection characteristics of the probes, and it is concluded...... that, for the imaging of optical-field intensity distributions containing predominantly evanescent-wave components, a sharp uncoated tip is the probe of choice. Complementary results obtained with optical phase-conjugation experiments with he uncoated probes are discussed in relation to the probe...

Developmental venous anomalies with capillary stain: a subgroup of symptomatic DVAs?

Energy Technology Data Exchange (ETDEWEB)

Roccatagliata, Luca [Hopital Foch, Service de Neuroradiologie Diagnostique et Therapeutique, Suresnes (France); University of Genoa, Department of Neurosciences, Ophthalmology and Genetics, Genoa (Italy); Berg, Rene van den [AMC, Department of Radiology, Amsterdam (Netherlands); Soderman, Michael [Karolinska University Hospital, Department of Neuroradiology, Stockholm (Sweden); Boulin, Anne; Condette-Auliac, Stephanie; Rodesch, Georges [Hopital Foch, Service de Neuroradiologie Diagnostique et Therapeutique, Suresnes (France)

2012-05-15

Intracranial developmental venous anomalies (DVAs) are considered benign vascular dispositions; they are asymptomatic in the vast majority of cases. They represent extreme variations of the venous drainage and may rarely be responsible for focal venous ischemia leading to neurological dysfunction. The aim of the study is to analyze a group of patients with symptomatic DVAs with capillary stain at angiography. We retrospectively reviewed the clinical and radiological features of patients in which a DVA was considered the cause of a neurological event. In all the patients, the DVA was suspected by angio-CT or MRI and conventional angiography was performed to detail the angioarchitecture of the DVA. A total of 7 patients and 11 DVAs were identified; three patients had multiple DVAs. Three DVAs were frontal, two were parietal, two were thalamic, one was in the midbrain, and three were cerebellar. Patients presented with progressive neurological deficits, seizures, or cerebral hemorrhage. All these DVAs were associated with a peculiar capillary stain at angiography. Although being normal anatomical variations, DVAs may create, because of hemodynamic unbalance, venous ischemia that induces angiogenic phenomena. MRI shows the suffering of the brain and angiography witnesses this angiogenesis under the form of capillary stain. Conventional angiography can thus provide useful information to recognize ''atypical'' symptomatic DVAs. (orig.)

Evaluation of the Gen-Probe DNA probe for the detection of legionellae in culture

International Nuclear Information System (INIS)

Edelstein, P.H.

1986-01-01

A commercial DNA probe kit designed to detect rRNA from legionellae was evaluated for its ability to correctly discriminate between legionellae and non-legionellae taken from culture plates. The probe kit, made by the Gen-Probe Corp. (San Diego, Calif.), was radiolabeled with 125 I, and probe bacterial RNA hybridization, detected in a simple one-tube system hybridization assay, was quantitated with a gamma counter. A total of 156 Legionella sp. strains were tested, of which 125 were Legionella pneumophila and the remainder were strains from 21 other Legionella spp. A total of 106 gram-negative non-legionellae, isolated from human respiratory tract (81%) and other body site (19%) specimens, were also tested; 14 genera and 28 species were represented. The probe easily distinguished all of the legionellae from the non-legionellae. The average legionellae/non-legionellae hybridization ratio was 42:1, and the lowest ratio was 2:1; a minor modification in the procedure increased the lowest ratio to 5:1. In addition to correctly identifying all Legionella species, the probe was able to separate some of the various species of Legionella. L. pneumophila strains hybridized more completely to the probe than did the other Legionella spp.; L. wadsworthii and L. oakridgensis hybridized only about 25% of the probe relative to L. pneumophila. Some strains of phenotypically identified L. pneumophila had much lower hybridization to the probe than other members of the species and may represent a new Legionella species. The simplicity of the technique and specificity of the probe make it a good candidate for confirming the identity of legionellae in culture

A hydrophobic organelle probe based on aggregation-induced emission: Nanosuspension preparation and direct use for endoplasmic reticulum imaging in living cells

Science.gov (United States)

Zheng, Sichao; Huang, Cuihong; Zhao, Xuyan; Zhang, Yong; Liu, Shuwen; Zhu, Qiuhua

2018-01-01

Organic fluorophores have a wide range of biological uses and are usually needed to be prepared as water-soluble compounds or nanoparticles for applications in aqueous biosystems owing to their hydrophobic properties, which often is a complex, time-consuming and high-cost process. Here, the nanoparticle preparation of hydrophobic fluorophores and their application in cell imaging have been investigated. It was found: a) fetal bovine serum (FBS) shows an excellent dispersion effect on hydrophobic small-molecule organic compounds; b) a hydrophobic C6-unsubstituted tetrahydropyrimidine (Me-THP-Naph) can be prepared as nanosuspensions utilizing cell culture medium with 10% FBS and directly be used as a specific real-time imaging probe for the endoplasmic reticulum (ER), a dynamic organelle playing a crucial role in many cellular processes. Compared with existing ER-targeted organic fluorescent probes, Me-THP-Naph, a product of an efficient five-component reaction that we developed, has unconventional aggregation-induced emission characteristics and shows advantages of low cost, long-term staining, good photostability, high signal-to-noise ratio and excellent biocompatibility, which make it a potential specific probe for real-time ER imaging. More importantly, this work affords a simple strategy for direct application of hydrophobic organic compounds in aqueous biological systems.

Where do pulse oximeter probes break?

Science.gov (United States)

Crede, S; Van der Merwe, G; Hutchinson, J; Woods, D; Karlen, W; Lawn, J

2014-06-01

Pulse oximetry, a non-invasive method for accurate assessment of blood oxygen saturation (SPO2), is an important monitoring tool in health care facilities. However, it is often not available in many low-resource settings, due to expense, overly sophisticated design, a lack of organised procurement systems and inadequate medical device management and maintenance structures. Furthermore medical devices are often fragile and not designed to withstand the conditions of low-resource settings. In order to design a probe, better suited to the needs of health care facilities in low-resource settings this study aimed to document the site and nature of pulse oximeter probe breakages in a range of different probe designs in a low to middle income country. A retrospective review of job cards relating to the assessment and repair of damaged or faulty pulse oximeter probes was conducted at a medical device repair company based in Cape Town, South Africa, specializing in pulse oximeter probe repairs. 1,840 job cards relating to the assessment and repair of pulse oximeter probes were reviewed. 60.2 % of probes sent for assessment were finger-clip probes. For all probes, excluding the neonatal wrap probes, the most common point of failure was the probe wiring (>50 %). The neonatal wrap most commonly failed at the strap (51.5 %). The total cost for quoting on the broken pulse oximeter probes and for the subsequent repair of devices, excluding replacement components, amounted to an estimated ZAR 738,810 (USD $98,508). Improving the probe wiring would increase the life span of pulse oximeter probes. Increasing the life span of probes will make pulse oximetry more affordable and accessible. This is of high priority in low-resource settings where frequent repair or replacement of probes is unaffordable or impossible.

Quantification of histochemical stains using whole slide imaging: development of a method and demonstration of its usefulness in laboratory quality control.

Science.gov (United States)

Gray, Allan; Wright, Alex; Jackson, Pete; Hale, Mike; Treanor, Darren

2015-03-01

Histochemical staining of tissue is a fundamental technique in tissue diagnosis and research, but it suffers from significant variability. Efforts to address this include laboratory quality controls and quality assurance schemes, but these rely on subjective interpretation of stain quality, are laborious and have low reproducibility. We aimed (1) to develop a method for histochemical stain quantification using whole slide imaging and image analysis and (2) to demonstrate its usefulness in measuring staining variation. A method to quantify the individual stain components of histochemical stains on virtual slides was developed. It was evaluated for repeatability and reproducibility, then applied to control sections of an appendix to quantify H&E staining (H/E intensities and H:E ratio) between automated staining machines and to measure differences between six regional diagnostic laboratories. The method was validated with laboratories from 0.57 to 0.89. A simple method using whole slide imaging can be used to quantify and compare histochemical staining. This method could be deployed in routine quality assurance and quality control. Work is needed on whole slide imaging devices to improve reproducibility. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

A double blind randomised controlled clinical trial comparing a novel anti-stain and calculus reducing dentifrice with a standard fluoride dentifrice.

Science.gov (United States)

Jowett, Adrian K; Marlow, Ian; Rawlinson, Andrew

2013-04-01

This clinical trial tested the anti-stain efficacy at 3 and 6 months of a novel, sodium polyaspartate-containing, anti-stain dentifrice. In addition, the efficacy of the new dentifrice in controlling gingival inflammation and inhibition of calculus deposition was tested. Participants were recruited to this double blind randomised control clinical trial, and allocated to either test or control groups. The presence of stain and calculus were entry criteria. Measurements of stain, calculus and gingival inflammation were recorded using the Shaw and Murray Stain score, Volpe-Manhold Calculus score and the Modified Gingival Index respectively. Measurements were made at baseline, prior to the removal of stain and calculus, and after 3 and 6 months. Missing data were imputed by and the outcomes were analysed using univariate analysis. At three months, toothpaste containing sodium polyaspartate was better (difference of mean 1.13 with SEM 0.57) than control for the control of dental stain (p0.05). There was no difference between toothpastes with respect to calculus deposition or gingival inflammation. Toothpaste containing sodium polyaspartate was more effective than a control toothpaste at preventing deposition of dental stain for 3 months after professional tooth cleaning but showed no significant effect at 6 months. Sodium polyaspartate toothpaste was more effective than a control toothpaste at preventing dental stain formation and maybe helpful in controlling staining between episodes of scaling and polishing. Copyright © 2012 Elsevier Ltd. All rights reserved.

Haematoxylin and eosin staining identifies medium to large bacterial aggregates with a reliable specificity: A comparative analysis of follicular bacterial aggregates in axillary biopsies using peptide nucleic acid-fluorescence in situ hybridization and haematoxylin and eosin staining.

Science.gov (United States)

Ring, Hans Christian; Theut Riis, Peter; Bay, Lene; Kallenbach, Klaus; Bjarnsholt, Thomas; Jemec, Gregor B E

2017-10-01

Although peptide nucleic acid (PNA), fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy (CLSM) are the reference tools in the study of bacterial aggregates/biofilms, it may also be rather time-consuming. This study aimed to investigate the sensitivity and specificity between bacterial aggregates identified by haematoxylin and eosin (HE) staining vs bacterial aggregates in corresponding PNA-FISH samples. Axillary biopsies were obtained in 24 healthy controls. HE-stained and PNA-FISH samples were investigated using traditional light microscopy and CLSM, respectively. The data demonstrate that HE staining identifies large bacterial aggregates (>10 μm) with a sensitivity of 0.43 and specificity of 1. The methods, however, are not equivalent as demonstrated by a McNemar's test (P=.04). Where bacterial aggregates >10 μm in diameter, HE staining may offer a rapid and practical low-cost tool to evaluate bacterial aggregates. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Color stability and staining of silorane after prolonged chemical challenges

DEFF Research Database (Denmark)

de Jesus, Vivian CBR; Martinelli, Nata Luiz; Poli-Frederico, Regina Célia

Objectives: The purpose of this study was to investigate the effect of prolonged chemical challenges on color stability and staining susceptibility of a silorane-based composite material when compared to methacrylate-based composites. Methods: Cylindrical specimens (n=24) were fabricated from...... methacrylate (Filtek Z250, 3M ESPE; Filtek Z350XT, 3M ESPE; Master Fill, Biodinâmica) or silorane-based (Filtek P90, 3M ESPE) composite materials. Initial color was registered in a spectrophotometer. Specimens were divided in four groups and individually stored at 37°C in 0.02N citric acid, 0.02N phosphoric...... acid, 75% ethanol or distilled water (control) for 7, 14, 21, and 180 days, when new measurements were performed. A staining test was performed (n=12) after 21 days of chemical challenge by immersion in coffee during 3 weeks at 37°C. Color changes (¿E) were characterized using the CIEL*a*b* color...

Adaptive segmentation of nuclei in H&S stained tendon microscopy

Science.gov (United States)

Chuang, Bo-I.; Wu, Po-Ting; Hsu, Jian-Han; Jou, I.-Ming; Su, Fong-Chin; Sun, Yung-Nien

2015-12-01

Tendiopathy is a popular clinical issue in recent years. In most cases like trigger finger or tennis elbow, the pathology change can be observed under H and E stained tendon microscopy. However, the qualitative analysis is too subjective and thus the results heavily depend on the observers. We develop an automatic segmentation procedure which segments and counts the nuclei in H and E stained tendon microscopy fast and precisely. This procedure first determines the complexity of images and then segments the nuclei from the image. For the complex images, the proposed method adopts sampling-based thresholding to segment the nuclei. While for the simple images, the Laplacian-based thresholding is employed to re-segment the nuclei more accurately. In the experiments, the proposed method is compared with the experts outlined results. The nuclei number of proposed method is closed to the experts counted, and the processing time of proposed method is much faster than the experts'.

NeuroMEMS: Neural Probe Microtechnologies

Directory of Open Access Journals (Sweden)

Sam Musallam

2008-10-01

Full Text Available Neural probe technologies have already had a significant positive effect on our understanding of the brain by revealing the functioning of networks of biological neurons. Probes are implanted in different areas of the brain to record and/or stimulate specific sites in the brain. Neural probes are currently used in many clinical settings for diagnosis of brain diseases such as seizers, epilepsy, migraine, Alzheimer’s, and dementia. We find these devices assisting paralyzed patients by allowing them to operate computers or robots using their neural activity. In recent years, probe technologies were assisted by rapid advancements in microfabrication and microelectronic technologies and thus are enabling highly functional and robust neural probes which are opening new and exciting avenues in neural sciences and brain machine interfaces. With a wide variety of probes that have been designed, fabricated, and tested to date, this review aims to provide an overview of the advances and recent progress in the microfabrication techniques of neural probes. In addition, we aim to highlight the challenges faced in developing and implementing ultralong multi-site recording probes that are needed to monitor neural activity from deeper regions in the brain. Finally, we review techniques that can improve the biocompatibility of the neural probes to minimize the immune response and encourage neural growth around the electrodes for long term implantation studies.

PHACE syndrome misdiagnosed as a port-wine stain

OpenAIRE

Thomson, Jason; Greig, Aina; Lloyd, Claire; Morrison, Danny; Flohr, Carsten

2015-01-01

We present the case of a boy born with a large macular, segmental vascular anomaly over the left face, initially diagnosed as a capillary malformation (port-wine stain) by the postnatal paediatric team. The vascular anomaly in the face then grew rapidly during the first few weeks of life and started to occlude the left eye, causing parental concerns about the infant's vision. A dermatological opinion established that the lesion was a segmental infantile haemangioma (IH). This, in combination ...

Fluorescent probes as a tool for labelling and tracking the amphibian chytrid fungus Batrachochytrium dendrobatidis.

Science.gov (United States)

Herbert, Sarah M; Leung, Tommy L F; Bishop, Phillip J

2011-09-09

The dissemination of the virulent pathogen Batrachochytrium dendrobatidis (Bd) has contributed to the decline and extinction of many amphibian species worldwide. Several different strains have been identified, some of which are sympatric. Interactions between co-infecting strains of a pathogen can have significant influences on disease epidemiology and evolution; therefore the dynamics of multi-strain infections is an important area of research. We stained Bd cells with 2 fluorescent BODIPY fatty acid probes to determine whether these can potentially be used to distinguish and track Bd cell lines in multi-strain experiments. Bd cells in broth culture were stained with 5 concentrations of green-fluorescent BODIPY FL and red-fluorescent BODIPY 558/568 and visualised under an epifluorescent microscope for up to 16 d post-dye. Dyed strains were also assessed for growth inhibition. The most effective concentration for both dyes was 10 pM. This concentration of dye produced strong fluorescence for 12 to 16 d in Bd cultures held at 23 degrees C (3 to 4 generations), and did not inhibit Bd growth. Cells dyed with BODIPY FL and BODIPY 558/568 can be distinguished from each other on the basis of their fluorescence characteristics. Therefore, it is likely that this technique will be useful for research into multi-strain dynamics of Bd infections.

Ulex Europaeus lectin and anti-CD31 staining in squamous cell carcinoma of the uterine cervix: potential prognostic markers.

Science.gov (United States)

Davidson, B; Goldberg, I; Gotlieb, W H; Lerner-Geva, L; Ben-Baruch, G; Kopolovic, J

1998-07-01

Seventy-five squamous cell carcinomas of the uterine cervix and 10 controls were stained for Ulex Europaeus lectin 1 (UEA-1) and anti-CD31, and the results were analyzed with respect to patient age, clinical stage, tumor grade, and survival during a follow-up period of 1 to 13 years. The patients' mean age at the time of diagnosis was 47.8 years (range, 27 to 83). Seventeen patients died of disease, 2 had disease recurrence, and 51 patients remained free of disease; 5 patients were lost to follow-up. Twenty-eight cases (37.3%) showed focal membranous staining for UEA-1 and 9 cases (12%) showed a diffuse pattern; 38 cases (50.7%) were UEA-1 negative. Poor survival was related to diffuse membranous UEA-1 immunoreactivity (p = 0.02), age (p = 0.014), grade (p = 0.02), and stage (p = 0.0002). CD31-positive neoplastic cells displayed a cytoplasmic pattern. Fifteen cases (20%) had diffuse staining and another 15 (20%) stained focally; 45 cases (60%) were CD31-negative. The adjacent nonneoplastic epithelium and all 10 controls were uniformly negative for CD31. Variable staining of the endocervical epithelium and weak or negative staining of ectocervical epithelium for UEA-1 were observed. However, the epithelium in all controls was negative for UEA-1. Poor survival was related to both focal and diffuse staining for CD31 (p = 0.01 and p = 0.03, respectively). Staining by both UEA-1 and anti-CD31 retained its correlation with survival after exclusion of stage la tumors.

Randomized controlled trial to evaluate tooth stain reduction with nicotine replacement gum during a smoking cessation program

LENUS (Irish Health Repository)

Whelton, Helen

2012-06-13

AbstractBackgroundIn addition to its general and periodontal health effects smoking causes tooth staining. Smoking cessation support interventions with an added stain removal or tooth whitening effect may increase motivation to quit smoking. Oral health professionals are well placed to provide smoking cessation advice and support to patients. The objective of the present study was to evaluate the effect of Nicorette® Freshmint Gum used in a smoking cessation programme administered in a dental setting, on extrinsic stain and tooth shade among smokers.MethodsAn evaluator-blinded, randomized, 12-week parallel-group controlled trial was conducted among 200 daily smokers motivated to quit smoking. Participants were randomised to use either the Nicorette® Freshmint Gum or Nicorette® Microtab (tablet). Tooth staining and shade were rated using the modified Lobene Stain Index and the Vita® Shade Guide at baseline, weeks 2, 6 and 12. To maintain consistency with other whitening studies, the primary end-point was the mean change in stain index between baseline and week 6. Secondary variables included changes in stain measurements and tooth shade at the other time points the number of gums or tablets used per day and throughout the trial period; and the number of cigarettes smoked per day. Treatments were compared using analysis of covariance (ANCOVA), using treatment and nicotine dependence as factors and the corresponding baseline measurement as a covariate. Each comparison (modified intention-to-treat) was tested at the 0.05 level, two-sided. Within-treatment changes from baseline were compared using a paired t-test.ResultsAt week 6, the gum-group experienced a reduction in mean stain scores whilst the tablet-group experienced an increase with mean changes of -0.14 and +0.12 respectively, (p = 0.005, ANCOVA). The change in mean tooth shade scores was statistically significantly greater in the gum-group than in the tablet group at 2 (p = 0.015), 6 (p = 0

Utility of Iron Staining in Identifying the Cause of Renal Allograft Dysfunction in Patients with Sickle Cell Disease

Directory of Open Access Journals (Sweden)

Yingchun Wang

2015-01-01

Full Text Available Sickle cell nephropathy (SCN is associated with iron/heme deposition in proximal renal tubules and related acute tubular injury (ATI. Here we report the utility of iron staining in differentiating causes of renal allograft dysfunction in patients with a history of sickle cell disease. Case 1: the patient developed acute allograft dysfunction two years after renal transplant. Her renal biopsy showed ATI, supported by patchy loss of brush border and positive staining of kidney injury molecule-1 in proximal tubular epithelial cells, where diffuse increase in iron staining (2+ was present. This indicated that ATI likely resulted from iron/heme toxicity to proximal tubules. Electron microscope confirmed aggregated sickle RBCs in glomeruli, indicating a recurrent SCN. Case 2: four years after renal transplant, the patient developed acute allograft dysfunction and became positive for serum donor-specific antibody. His renal biopsy revealed thrombotic microangiopathy (TMA and diffuse positive C4d stain in peritubular capillaries. Iron staining was negative in the renal tubules, implying that TMA was likely associated with acute antibody-mediated rejection (AAMR, type 2 rather than recurrent SCN. These case reports imply that iron staining is an inexpensive but effective method in distinguishing SCN-associated renal injury in allograft kidney from other etiologies.

Effect of finishing and polishing on the color stability of a composite resin immersed in staining solutions

Directory of Open Access Journals (Sweden)

Maiara Justo Polli

2015-01-01

Full Text Available Objective: To evaluate the influence of finishing/polishing methods and staining solutions using different immersion periods on the color stability of a microhybrid composite resin. Materials and Methods: Ninety specimens were fabricated using a stainless steel mold and polyester strips. The samples were randomly divided into five groups according to the finishing and polishing performed: Control group (no surface treatment was performed, Diamond Pro group, Diamond burs group, Enhance group, and SiC paper group. After finishing and polishing, six samples from each group were immersed in coffee, red wine, or water for 30 days. The color measurements were obtained using digital photography before immersion and after 7, 15, and 30 days of immersion. The red, green, and blue values provided by the Adobe Photoshop software were converted into CIELab values. A three-way analysis of variance and Tukey's test were used for statistical analysis (P ≤ 0.05. Results: The finishing and polishing methods, staining solutions, immersion times, and their interaction had statistically significant effects on the color change (P = 0.00. Coffee and red wine caused intense staining. Among the polishing methods, the highest color change value was observed in the control group (P < 0.05 and the Diamond Pro disks provided the most stain-resistant surfaces (P ≤ 0.05. Conclusion: The finishing and polishing method, staining solution, and immersion time influences the color stability. Finishing and polishing should be applied to obtain a more stain-resistant surface.

Double epi-illumination microscopy with separate visualization of two antigens: a combination of epi-polarization for immunogold-silver staining and epi-fluorescence for alkaline phosphatase staining

NARCIS (Netherlands)

van der Loos, C. M.; Becker, A. E.

1994-01-01

We present a method for an epi-illumination immunohistochemical double staining approach. The method combines the use of an immuno-alkaline phosphatase technique and the immunogold-silver technique, visualized with epifluorescence and epi-polarization illumination, respectively. Out of six tested

High-performance and anti-stain coating for porcelain stoneware tiles based on nanostructured zirconium compounds.

Science.gov (United States)

Ambrosi, Moira; Santoni, Sergio; Giorgi, Rodorico; Fratini, Emiliano; Toccafondi, Nicola; Baglioni, Piero

2014-10-15

The technological characteristics of porcelain stoneware tiles make them suitable for a wide range of applications spanning far beyond traditional uses. Due to the high density, porcelain stoneware tiles show high bending strength, wear resistance, surface hardness, and high fracture toughness. Nevertheless, despite being usually claimed as stain resistant, the surface porosity renders porcelain stoneware tiles vulnerable to dirt penetration with the formation of stains that can be very difficult to remove. In the present work, we report an innovative and versatile method to realize stain resistant porcelain stoneware tiles. The tile surface is treated by mixtures of nanosized zirconium hydroxide and nano- and micron-sized glass frits that thanks to the low particle dimension are able to penetrate inside the surface pores. The firing step leads to the formation of a glass matrix that can partially or totally close the surface porosity. As a result, the fired tiles become permanently stain resistant still preserving the original esthetical qualities of the original material. Treated tiles also show a remarkably enhanced hardness due to the inclusion of zirconium compounds in the glass coating. Copyright © 2014 Elsevier Inc. All rights reserved.

Impact of changing from staining to culture techniques on detection rates of Campylobacter spp. in routine stool samples in Chile.

Science.gov (United States)

Porte, Lorena; Varela, Carmen; Haecker, Thomas; Morales, Sara; Weitzel, Thomas

2016-05-13

Campylobacter is a leading cause of bacterial gastroenteritis, but sensitive diagnostic methods such as culture are expensive and often not available in resource limited settings. Therefore, direct staining techniques have been developed as a practical and economical alternative. We analyzed the impact of replacing Campylobacter staining with culture for routine stool examinations in a private hospital in Chile. From January to April 2014, a total of 750 consecutive stool samples were examined in parallel by Hucker stain and Campylobacter culture. Isolation rates of Campylobacter were determined and the performance of staining was evaluated against culture as the gold standard. Besides, isolation rates of Campylobacter and other enteric pathogens were compared to those of past years. Campylobacter was isolated by culture in 46 of 750 (6.1 %) stool samples. Direct staining only identified three samples as Campylobacter positive and reached sensitivity and specificity values of 6.5 and 100 %, respectively. In comparison to staining-based detection rates of previous years, we observed a significant increase of Campylobacter cases in our patients. Direct staining technique for Campylobacter had a very low sensitivity compared to culture. Staining methods might lead to a high rate of false negative results and an underestimation of the importance of campylobacteriosis. With the inclusion of Campylobacter culture, this pathogen became a leading cause of intestinal infection in our patient population.

Amalgam stained dentin: a proper substrate for bonding resin composite?

NARCIS (Netherlands)

Scholtanus, J.D.

2016-01-01

Nowadays the use of dental amalgam is mostly abandoned and substituted by tooth colored resin composites that can be bonded to teeth tissues by adhesive techniques. The aim of this thesis was to find out whether dark stained dentin, as often observed after removal of amalgam restorations and

Initial data release of regular blood drip stain created by varying fall height, angle of impact and source dimension

Directory of Open Access Journals (Sweden)

Nabanita Basu

2016-09-01

Full Text Available The dataset developed consists of 108 blood drip stains developed with fresh porcine blood, blood admixed with different dosage of Warfarin and Heparin, respectively. For each particular blood type (i.e. fresh blood, blood admixed with Warfarin at different dosage and blood admixed with Heparin at varied dosage stain patterns were created by passive dripping of blood from a 2.5 cm3 subcutaneous syringe with needle filled to capacity, at 30°, 60° and 90° angle of impact with corresponding fall height of 20, 40 and 60 cm respectively. In the other dataset of 162 datapoints, 81 regular drip stains were formed from blood that had dripped passively from a subcutaneous syringe without needle at the aforementioned angle of impact and fall height, while the other stains were formed as a result of dripping of blood from a subcutaneous syringe with needle. In order to compare stains formed, all stains were recorded on the same representative, non-porous, smooth target surface under similar physical conditions. The interpretations relevant to the dataset are available in the article titled ‘2D Source Area prediction based on physical characteristics of a regular, passive blood drip stain’ (Basu and Bandyopadhyay, 2016 [7]. An image pre-processing algorithm for extracting ROI has also been incorporated in this article. Keywords: Drip stain, Bloodstain Pattern Analysis, Source Dimension prediction

Preventing probe induced topography correlated artifacts in Kelvin Probe Force Microscopy

NARCIS (Netherlands)

Polak, L.; Wijngaarden, Rinke J.

2016-01-01

Kelvin Probe Force Microscopy (KPFM) on samples with rough surface topography can be hindered by topography correlated artifacts. We show that, with the proper experimental configuration and using homogeneously metal coated probes, we are able to obtain amplitude modulation (AM) KPFM results on a

A novel hematoxylin and eosin stain assay for detection of the parasitic dinoflagellate Amoebophrya.

Science.gov (United States)

Li, Caiwen; Chen, Tiantian

2017-02-01

The parasitic dinoflagellate Amoebophrya infects broad range of marine organisms. Particularly, Amoebophrya infections in planktonic dinoflagellates can prevent or delay the formation of algal blooms, and recycle undergrazed planktonic dinoflagellates back to the microbial loop by disrupting host cells. Its ecological significance was gradually recognized along with the discovery of its enormous molecular diversity in oceanic and coastal ecosystems. Thus, we developed a reliable, easily accessible and less time-consuming assay, to detect and assess Amoebophrya infections in planktonic dinoflagellates. The modified hematoxylin and eosin staining assay provided reliable diagnosis of Amoebophrya infection by identifying the characteristic "beehive" of the multinucleate trophonts. After staining, the typical multinucleate "beehive" is evidently distinguishable from the compact nuclei of uninfected host cells. The modified hematoxylin and eosin (H & E) staining assay is easy to use, that can be routinely performed within 3h (up to 20 samples/batch) using general laboratory equipment, supplies and chemical reagents. The produced slides with agar-embedded dinoflagellate cells can be stored for several months or even years in a dry place without noticeable loss in quality of staining. With suitable calculation, the modified H & E assay can be applied to assess the prevalence of Amoebophrya infection in planktonic dinoflagellates. This efficient and powerful assay will facilitate the investigation on the ecological roles of Amoebophryidae in coastal and oceanic ecosystem. Copyright © 2016 Elsevier B.V. All rights reserved.

Autometallographic (AMG) technique used for enhancement of the Golgi-Cox staining gives good contrast andhigh resolution of dendrites and spines

DEFF Research Database (Denmark)

Orlowski, Dariusz

Despite the existence of many newer staining methods, Golgi staining still remains the primary method forvisualization of the dendrites and spines. The black deposit in the Golgi-Cox impregnated cells is a Mercuricsulphide, therefore autometallographic (AMG) technique which is used for visualizat...... of dendrites and spines in the rat hippocampus. The describedmethod will be of value for future behavioural-anatomical studies, examining changes in dendrite branching andspine density caused by brain diseases and their subsequent treatment.......Despite the existence of many newer staining methods, Golgi staining still remains the primary method forvisualization of the dendrites and spines. The black deposit in the Golgi-Cox impregnated cells is a Mercuricsulphide, therefore autometallographic (AMG) technique which is used...... for visualization of the metals and metalsulphides/selenides in tissue may be used to enhance the Golgi-Cox staining. We demonstrated accordingly thatuse of AMG enhancement method on the Golgi-Cox staining gives good contrast and high resolution of dendritesand spines. Moreover, this method is cheaper and more...

Accuracy of micro four-point probe measurements on inhomogeneous samples: A probe spacing dependence study

DEFF Research Database (Denmark)

Wang, Fei; Petersen, Dirch Hjorth; Østerberg, Frederik Westergaard

2009-01-01

In this paper, we discuss a probe spacing dependence study in order to estimate the accuracy of micro four-point probe measurements on inhomogeneous samples. Based on sensitivity calculations, both sheet resistance and Hall effect measurements are studied for samples (e.g. laser annealed samples...... the probe spacing is smaller than 1/40 of the variation wavelength, micro four-point probes can provide an accurate record of local properties with less than 1% measurement error. All the calculations agree well with previous experimental results.......) with periodic variations of sheet resistance, sheet carrier density, and carrier mobility. With a variation wavelength of ¿, probe spacings from 0.0012 to 1002 have been applied to characterize the local variations. The calculations show that the measurement error is highly dependent on the probe spacing. When...

Eddy-current probe design

International Nuclear Information System (INIS)

Kincaid, T.G.; McCary, R.O.

1983-01-01

This paper describes theoretical and experimental work directed toward finding the optimum probe dimensions and operating frequency for eddy current detection of half-penny surface cracks in nonmagnetic conducting materials. The study applies to probes which excite an approximately uniform spatial field over the length of the crack at the surface of the material. In practical terms, this means that the probe is not smaller than the crack length in any of its critical dimensions. The optimization of a simple coil probe is first analyzed in detail. It is shown that signal-to-noise ratio and lift-off discrimination are maximized by a pancake coil with mean radius not greater than the crack length, operated at a frequency which gives a skin depth equal to the crack depth. The results obtained for the simple coil are then used as a basis for discussion of the design of coils with ferrite cores and shields, and for the design of recording head type probes

Comparison of three staining methods for the detection of intestinal microspora spp.

Directory of Open Access Journals (Sweden)

Khadijeh Khanaliha

2014-12-01

Full Text Available This study aimed to compare three staining methods including: Calcofluor white, Chromotrope and Quick Hot Gram chromotrope used in diagnosis of intestinal microsporidial spores.One hundred and seventy five stool specimens were collected from patients referred to Laboratory of Intestinal Protozoology at the School of Public Health, Tehran University of Medical Sciences during 2012-2013. All of specimens were evaluated by nested PCR. The formalin-fixed stool samples were prepared from each specimen and dried at room temperature for 10 min, followed by 10 min methanol fixation. All the collected stool samples were evaluated blindly by calcofluor white, Chromotrope and Quick Hot Gram chromotrope staining methods separately.Microsporidial spores were recognized using Chromotrope, Quick Hot Gram chromotrope and Calcofluor white, in16 of 18 (88.8%, 17 of 18 (94.4% and 18 of 18(100% samples that were positive by nested PCR respectively. Regarding 14 stool samples that were negative by nested PCR, 14 cases were negative by chromotrope and Quick hot Gram chromotrope and 13 samples were negative by Calcofluor white. One discordant sample interpreted as false positive.Calcofluor white staining had the best performance for the detection of intestinal Microsprora spores and can be used as initial screen test for the detection of intestinal Microspora spp.

New visible and selective DNA staining method in gels with tetrazolium salts.

Science.gov (United States)

Paredes, Aaron J; Naranjo-Palma, Tatiana; Alfaro-Valdés, Hilda M; Barriga, Andrés; Babul, Jorge; Wilson, Christian A M

2017-01-15

DNA staining in gels has historically been carried out using silver staining and fluorescent dyes like ethidium bromide and SYBR Green I (SGI). Using fluorescent dyes allows recovery of the analyte, but requires instruments such as a transilluminator or fluorimeter to visualize the DNA. Here we described a new and simple method that allows DNA visualization to the naked eye by generating a colored precipitate. It works by soaking the acrylamide or agarose DNA gel in SGI and nitro blue tetrazolium (NBT) solution that, when exposed to sunlight, produces a purple insoluble formazan precipitate that remains in the gel after exposure to light. A calibration curve made with a DNA standard established a detection limit of approximately 180 pg/band at 500 bp. Selectivity of this assay was determined using different biomolecules, demonstrating a high selectivity for DNA. Integrity and functionality of the DNA recovered from gels was determined by enzymatic cutting with a restriction enzyme and by transforming competent cells after the different staining methods, respectively. Our method showed the best performance among the dyes employed. Based on its specificity, low cost and its adequacy for field work, this new methodology has enormous potential benefits to research and industry. Copyright © 2016 Elsevier Inc. All rights reserved.

Clinical outcomes of double staining and additional ILM peeling during ERM surgery.

Science.gov (United States)

Oh, Ha Na; Lee, Joo Eun; Kim, Hyun Woong; Yun, Il Han

2013-08-01

To assess the clinical outcomes in idiopathic epiretinal membrane (ERM) patients after vitrectomy and ERM removal with or without additional indocyanine green (ICG)-assisted internal limiting membrane (ILM) peeling. The medical records of 43 patients with an idiopathic ERM that underwent vitrectomy and ERM removal between July 2007 and April 2010 were reviewed. The patients were divided into two groups: triamcinolone-assisted simple ERM peeling only (group A, n = 23) and triamcinolone-assisted ERM peeling followed by ICG staining and peeling of the remaining internal ILM (group B, n = 20). No difference was found between the two groups in terms of visual acuity, macular thickness, P1 amplitude or implicit time on multifocal-electroretinogram (mfERG) at six and 12 months postoperatively. In group B, ICG staining after ERM peeling demonstrated that the ILM had been removed together with the ERM in 12 eyes (60%), and all 12 eyes showed punctate retinal hemorrhages during ERM peeling. There was no recurrence of an ERM in either group. Additional procedures involving ICG staining and ILM peeling during ERM surgery do not appear to have an additive effect on the clinical outcomes in terms of visual acuity, retinal function based on mfERG, or recurrence rate.

Wearable probes for service design

DEFF Research Database (Denmark)

Mullane, Aaron; Laaksolahti, Jarmo Matti; Svanæs, Dag

2014-01-01

Probes are used as a design method in user-centred design to allow end-users to inform design by collecting data from their lives. Probes are potentially useful in service innovation, but current probing methods require users to interrupt their activity and are consequently not ideal for use...... by service employees in reflecting on the delivery of a service. In this paper, we present the ‘wearable probe’, a probe concept that captures sensor data without distracting service employees. Data captured by the probe can be used by the service employees to reflect and co-reflect on the service journey......, helping to identify opportunities for service evolution and innovation....

The incidence of IgG4-positive plasma cells staining TIN in patients with biopsy-proven tubulointerstitial nephritis.

Science.gov (United States)

Mac, Kathy; Wu, Xiao Juan; Mai, Jun; Howlin, Kenneth; Suranyi, Michael; Yong, Jim; Makris, Angela

2017-06-01

IgG4 disease is rare. However, IgG4 tubulointerstitial nephritis (TIN) is the most common renal manifestation. IgG4 disease is usually associated with elevated serum IgG4 levels and other organ involvement, low-density renal lesions on enhanced CT imaging and immune activation. The incidence of IgG4-TIN may be underestimated, as staining for IgG4 is not routine. This study sought to describe the prevalence of previously undiagnosed IgG4-TIN. Due to the complexity of the diagnosis, we only attempt to look at IgG4-positive plasma cell TIN as a potential indication for IgG4 renal disease. A retrospective review of native renal biopsies performed between 2002 and 2012 with a primary diagnosis of TIN was selected. Samples for which interstitial nephritis was secondary to a glomerular disease were excluded. The tissues were stained for IgG4 and scored by two blinded observers. Demographic and follow-up details were collected. This study was approved by the local ethics committee. 82 cases of interstitial nephritis from a total of 1238 renal biopsies (2002-2012) were available after staining for further assessment. 12 samples demonstrated staining consistent with the criteria for IgG4-positive plasma cell TIN, of which 3 had mildly positive staining, 7 moderately positive staining and 2 had markedly positive staining. There were no statistically significant differences in the baseline characteristics between the positive and negative staining groups. A number of cases of IgG4-positive plasma cell TIN were observed histologically that had been previously diagnosed as non-specific chronic TIN. IgG4-positive plasma cell TIN made up 1% of all renal biopsies performed over 10 years and 13% of all biopsies demonstrating TIN not related to glomerular disease. IgG4 staining should be considered routinely in biopsies demonstrating primary TIN. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

Effect of amnioinfusion for meconium stained amniotic fluid on perinatal outcome.

Science.gov (United States)

Ashfaq, F; Shah, A A

2004-06-01

To see the effect of amnioinfusion on perinatal outcome in cases of meconium staining of liquor. This study was conducted in department of Obstetrics and Gynaecology, unit 1, Jinnah Postgraduate Medical Centre, Karachi, from 1st January 1998 to 31st December 2000. Four hundred patients were included in this study, assigning 200 for amnioinfusion and 200 as control. All patients were matched in both the groups with respect to age, antenatal booking, parity, gestational age, stage of labour, colour of amniotic fluid and fetal birth weight. Both the groups were found to be comparable. The rate of Caesarean section was found to be 37% in amnioinfusion group, which collaborates with other international studies. The fetal outcome was better i.e. 91% alive and healthy, after amnioinfusion due to dilution of meconium stained amniotic fluid with physiological solutions. The perinatal outcome was recorded by Apgar score at 5 minutes. The perinatal morbidity and mortality both were significantly lowered and was found to be 6% as compared to 14% in control, which was also noticed by less number of admissions in nursery i.e. 12% and perinatal deaths. The incidence of meconium aspiration syndrome was found to be 56% in control and was reduced to 22% after amnioinfusion in the other arm of the study. These results are very encouraging and suggestion can be safely made that in future amnioinfusion will be the ideal method of preventing fetal distress due to meconium stained amniotic fluid.

Aspheric surface measurement using capacitive probes

Science.gov (United States)

Tao, Xin; Yuan, Daocheng; Li, Shaobo

2017-02-01

With the application of aspheres in optical fields, high precision and high efficiency aspheric surface metrology becomes a hot research topic. We describe a novel method of non-contact measurement of aspheric surface with capacitive probe. Taking an eccentric spherical surface as the object of study, the averaging effect of capacitive probe measurement and the influence of tilting the capacitive probe on the measurement results are investigated. By comparing measurement results from simultaneous measurement of the capacitive probe and contact probe of roundness instrument, this paper indicates the feasibility of using capacitive probes to test aspheric surface and proposes the compensation method of measurement error caused by averaging effect and the tilting of the capacitive probe.

Background-free, high sensitivity staining of proteins in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gels using a luminescent ruthenium complex.

Science.gov (United States)

Berggren, K; Chernokalskaya, E; Steinberg, T H; Kemper, C; Lopez, M F; Diwu, Z; Haugland, R P; Patton, W F

2000-07-01

SYPRO Ruby dye is a permanent stain comprised of ruthenium as part of an organic complex that interacts noncovalently with proteins. SYPRO Ruby Protein Gel Stain provides a sensitive, gentle, fluorescence-based method for detecting proteins in one-dimensional and two-dimensional sodium dodecyl sulfate-polyacrylamide gels. Proteins are fixed, stained from 3h to overnight and then rinsed in deionized water or dilute methanol/acetic acid solution for 30 min. The stain can be visualized using a wide range of excitation sources commonly used in image analysis systems including a 302 nm UV-B transilluminator, 473 nm second harmonic generation (SHG) laser, 488 nm argon-ion laser, 532 nm yttrium-aluminum-garnet (YAG) laser, xenon arc lamp, blue fluorescent light bulb or blue light-emitting diode (LED). The sensitivity of SYPRO Ruby Protein Gel Stain is superior to colloidal Coomassie Brilliant Blue (CBB) stain or monobromobimane labeling and comparable with the highest sensitivity silver or zinc-imidazole staining procedures available. The linear dynamic range of SYPRO Ruby Protein Gel stain extends over three orders of magnitude, which is vastly superior to silver, zinc-imidazole, monobromobimane and CBB stain. The fluorescent stain does not contain superfluous chemicals (formaldehyde, glutaraldehyde, Tween-20) that frequently interfere with peptide identification in mass spectrometry. While peptide mass profiles are severely altered in protein samples prelabeled with monobromobimane, successful identification of proteins by peptide mass profiling using matrix-assisted laser desorption/ionization mass spectrometry was easily performed after protein detection with SYPRO Ruby Protein Gel stain.

Solid-Color Stains on Western Redcedar and Redwood Siding

Science.gov (United States)

Mark Knaebe

2013-01-01

You have decided to put wood siding on your new house. Several questions are probably going through your mind: “What’s the best type of wood?” “Should I use paint or stain?” “Should I apply the finish before or after I install the siding?”

ANALISIS POTENSI PENERIMAAN KUALITAS ALUMNI PROGRAM STUDI EKONOMI SYARI’AH STAIN KUDUS DITINJAU DARI PERSPEKTIF STAKEHOLDER

Directory of Open Access Journals (Sweden)

Ekawati Rahayu Ningsih

2015-03-01

Full Text Available GRADUATES RECEPTION QUALITYANALYSIS OF THE POTENTIAL ECONOMIC STUDIES SYARI’AH IN STAIN KUDUS VIEWED FROM THE PERSPECTIVE STAKEHOLDER. The first aim of  this study was to determine the potential reception of  graduates quality in Shariah Economic Studiesof  STAINKudus in the world of  work. Second, to determine the motivations and needs of  stakeholders on the acceptance of  the quality of  graduates in Economics   Shariah STAIN Kudus.And third, to determine what factors which are supporting and inhibiting the absorption of graduates of the Department of Shariah Economics STAIN Kudus in working world. The theory that was developed as a basis for the analysis is the pyramid theory of  motivation and needs of  Abraham Maslow. By using qualitative research approach, the analysis and discussion of this study are:First, the potential acceptance of  Shariah Economy graduates in the working world, especially in the banking and financial institutions Shari’ah is still very large and potentially growing along with the rapid growth in the number of  banking and financial institutions Shari’ah in Indonesia. Second, motivation and needs of  stakeholders for the graduates reception of Shariah Economic STAIN Kudus is because it is the only college in the state of  religion around the Pantura area having Shariah Economic Studies Program and easily accessible. In addition, in order to establish a more synergistic relationship with the STAIN kudus then either the shari’a banking and financial institutions are willing to accept graduates of  Shariah Economic Studies Program as employees, of course, with the various criteria established in the job requirements. Keywords:Potential,  Quality  Admissions,  Graduates, Economic Shariah, Stakeholder. Tujuan penelitian ini adalah: Pertama, untuk mengetahui potensi penerimaan kualitas alumni Program Studi Ekonomi Syari’ah STAIN Kudus di dunia kerja. Kedua, untuk mengetahui motivasi dan

Effect of bleaching and repolishing procedures on coffee and tea stain removal from three anterior composite veneering materials.

Science.gov (United States)

Türkün, L Sebnem; Türkün, Murat

2004-01-01

Discolored teeth can be treated with resin veneers, but their color changes when confronted with staining solutions. Polishing procedures can provide a remedy for highly stained composites, but they tend to remove some materials as well. However, bleaching procedures are an effective, nondestructive method for solving the problem. The aim of this study was to compare the color change of three veneer composites exposed to staining solutions and to evaluate the effectiveness of a 15% hydrogen peroxide bleaching agent and three polishing systems to remove the stain. Forty-five disks (12 x 2 mm) each of Clearfil ST (Kuraray Co. Ltd., Osaka, Japan), Esthet-X (Dentsply/Caulk, Milford DE, USA), and Filtek A110 (3M ESPE, St. Paul, MN, USA) were prepared. The specimens were polished with Sof-Lex (3M ESPE), Enhance (Dentsply/Caulk), or PoGo (Dentsply/Caulk). Five specimens for each material-polishing system combination were immersed in coffee (Nescafe Classic, Nestle SA, Vevey, Switzerland) or tea (Earl Grey, Lipton, Blackfriars-London, England) for 7 days. The remaining disks were stored in water. Color measurements were made with a spectrophotometer (X-Rite Seroice SP78, Loaner, Köln, Germany) at baseline; after 1, 3, 5, and 7 days; and after bleaching and repolishing. After 1 week, one side of the specimens was bleached with Illuminé-office (Dentsply De Trey GmbH, Konstanz, Germany) for 1 hour, and the other side was repolished for 30 seconds. All comparisons of color change for the polishing systems, times, and staining solutions were subjected to repeated measurements of analysis of variance. Paired t-test was used to examine whether significant color differences (deltaE*) occurred during immersion at the specified time intervals (p < or = .05). Filtek A110 was the least stained resin composite. Its color remained under a deltaE* value of 2 during the study. Clearfil ST exhibited the most color change after 1 week. All specimens polished with Enhance showed less

A Simple and Efficient Method for Preparing Cell Slides and Staining without Using Cytocentrifuge and Cytoclips

Directory of Open Access Journals (Sweden)

Xiaotang Hu

2015-01-01

Full Text Available Cell staining is a necessary and useful technique for visualizing cell morphology and structure under a microscope. This technique has been used in many areas such as cytology, hematology, oncology, histology, virology, serology, microbiology, cell biology, and immunochemistry. One of the key pieces of equipment for preparing a slide for cell staining is cytology centrifuge (cytocentrifuge such as cytospin. However, many small labs do not have this expensive equipment and its accessory, cytoclips (also expensive relatively, which makes them difficult to study cell cytology. Here we present an alternative method for preparing a slide and cell staining in the absence of a cytocentrifuge (and cytoclips. This method is based on the principle that a regular cell centrifuge can be used to concentrate cells harvested from cell culture and then deposit the concentrated cell suspension to a slide evenly by using a cell spreader, followed by cell staining. The method presented is simple, rapid, economic, and efficient. This method may also avoid a possible change in cell morphology induced by cytocentrifuge.

Cantilevered probe detector with piezoelectric element

Science.gov (United States)

Adams, Jesse D; Sulchek, Todd A; Feigin, Stuart C

2013-04-30

A disclosed chemical detection system for detecting a target material, such as an explosive material, can include a cantilevered probe, a probe heater coupled to the cantilevered probe, and a piezoelectric element disposed on the cantilevered probe. The piezoelectric element can be configured as a detector and/or an actuator. Detection can include, for example, detecting a movement of the cantilevered probe or a property of the cantilevered probe. The movement or a change in the property of the cantilevered probe can occur, for example, by adsorption of the target material, desorption of the target material, reaction of the target material and/or phase change of the target material. Examples of detectable movements and properties include temperature shifts, impedance shifts, and resonant frequency shifts of the cantilevered probe. The overall chemical detection system can be incorporated, for example, into a handheld explosive material detection system.

The time domain triple probe method

International Nuclear Information System (INIS)

Meier, M.A.; Hallock, G.A.; Tsui, H.Y.W.; Bengtson, R.D.

1994-01-01

A new Langmuir probe technique based on the triple probe method is being developed to provide simultaneous measurement of plasma temperature, potential, and density with the temporal and spatial resolution required to accurately characterize plasma turbulence. When the conventional triple probe method is used in an inhomogeneous plasma, local differences in the plasma measured at each probe introduce significant error in the estimation of turbulence parameters. The Time Domain Triple Probe method (TDTP) uses high speed switching of Langmuir probe potential, rather than spatially separated probes, to gather the triple probe information thus avoiding these errors. Analysis indicates that plasma response times and recent electronics technology meet the requirements to implement the TDTP method. Data reduction techniques of TDTP data are to include linear and higher order correlation analysis to estimate fluctuation induced particle and thermal transport, as well as energy relationships between temperature, density, and potential fluctuations

Dithizone staining of intracellular zinc: an unexpected and versatile counterscreen for auxotrophic marker genes in Saccharomyces cerevisiae.

Directory of Open Access Journals (Sweden)

Daniel S Yuan

Full Text Available Auxotrophic marker genes such as URA3, LEU2, and HIS3 in Saccharomyces cerevisiae have long been used to select cells that have been successfully transformed with recombinant DNA. A longstanding challenge in working with these genes is that counterselection procedures are often lacking. This paper describes the unexpected discovery of a simple plate assay that imparts a bright red stain to cells experiencing nutritional stress from the lack of a marker gene. The procedure specifically stains a zinc-rich vesicular compartment analogous to the zinc-rich secretory vesicles found in insulin-secreting pancreatic islet cells and glutamate-secreting neurons. Staining was greatly diminished in zap1 mutants, which lack a homeostatic activator of zinc uptake, and in cot1 zrc1 double mutants, which lack the two yeast homologs of mammalian vesicle-specific zinc export proteins. Only one of 93 strains with temperature-sensitive alleles of essential genes exhibited an increase in dithizone staining at its non-permissive temperature, indicating that staining is not simply a sign of growth-arrested or dying cells. Remarkably, the procedure works with most commonly used marker genes, highlights subtle defects, uses no reporter constructs or expensive reagents, requires only a few hours of incubation, yields visually striking results without any instrumentation, and is not toxic to the cells. Many potential applications exist for dithizone staining, both as a versatile counterscreen for auxotrophic marker genes and as a powerful new tool for the genetic analysis of a biomedically important vesicular organelle.