High positive predictive value of Gram stain on catheter-drawn blood samples for the diagnosis of catheter-related bloodstream infection in intensive care neonates.

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Deleers, M; Dodémont, M; Van Overmeire, B; Hennequin, Y; Vermeylen, D; Roisin, S; Denis, O

2016-04-01

Catheter-related bloodstream infections (CRBSIs) remain a leading cause of healthcare-associated infections in preterm infants. Rapid and accurate methods for the diagnosis of CRBSIs are needed in order to implement timely and appropriate treatment. A retrospective study was conducted during a 7-year period (2005-2012) in the neonatal intensive care unit of the University Hospital Erasme to assess the value of Gram stain on catheter-drawn blood samples (CDBS) to predict CRBSIs. Both peripheral samples and CDBS were obtained from neonates with clinically suspected CRBSI. Gram stain, automated culture and quantitative cultures on blood agar plates were performed for each sample. The paired quantitative blood culture was used as the standard to define CRBSI. Out of 397 episodes of suspected CRBSIs, 35 were confirmed by a positive ratio of quantitative culture (>5) or a colony count of CDBS culture >100 colony-forming units (CFU)/mL. All but two of the 30 patients who had a CDBS with a positive Gram stain were confirmed as having a CRBSI. Seven patients who had a CDBS with a negative Gram stain were diagnosed as CRBSI. The sensitivity, specificity, positive predictive value and negative predictive value of Gram stain on CDBS were 80, 99.4, 93.3 and 98.1 %, respectively. Gram staining on CDBS is a viable method for rapidly (<1 h) detecting CRBSI without catheter withdrawal.

Techniques for controlling variability in gram staining of obligate anaerobes.

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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

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

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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

Lipid-rich carcinoma of the breast that is strongly positive for estrogen receptor: a case report and literature review

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Oba T

2016-03-01

Full Text Available Takaaki Oba,1 Mayu Ono,1 Asumi Iesato,1 Toru Hanamura,1 Takayuki Watanabe,1 Tokiko Ito,1 Toshiharu Kanai,1 Kazuma Maeno,1 Ken-ichi Ito,1 Ayako Tateishi,2 Akihiko Yoshizawa,2 Fumiyoshi Takayama31Division of Breast, Endocrine and Respiratory Surgery, Department of Surgery, Shinshu University School of Medicine, Matsumoto, Nagano, 2Department of Laboratory Medicine, Shinshu University Hospital, 3Imaging Center, Ichinose Neurosurgical Hospital, Matsumoto, JapanAbstract: Lipid-rich carcinoma (LRC of the breast is a rare breast cancer variant that accounts for <1% of all breast malignancies. It has been reported that LRCs are negative for estrogen receptor. Here, we report a case of LRC of the breast that was strongly positive for estrogen receptor and treated with endocrine adjuvant therapy. A 52-year-old postmenopausal female noticed a lump in her right breast by self-examination and presented to our hospital. Physical examination revealed an elastic 30 mm ×20 mm hard mass in the upper medial part of her right breast. The findings obtained using ultrasonography, mammography, and contrast-enhanced magnetic resonance imaging suggested breast cancer. Core needle biopsy resulted in the diagnosis of invasive carcinoma. The patient underwent mastectomy and sentinel lymph node biopsy. Histopathologically, the tumor cells were abundant in foamy cytoplasm. Because the presence of marked cytoplasmic lipid droplets was confirmed by Sudan IV staining and electron microscopic examination of the tumor and the lipid droplets were negative for periodic acid–Schiff staining, the tumor was diagnosed as an LRC. Immunohistochemically, estrogen and progesterone receptors of the tumor were strongly positive, human epidermal growth factor receptor type 2 was negative, and the ratio of Ki-67-positive cells was ~30%. After surgery, the patient underwent combination chemotherapy with anthracycline, cyclophosphamide, and 5-fluorouracil, followed by docetaxel. Thereafter

Diagnostic utility of melanin production by fungi: Study on tissue sections and culture smears with Masson-Fontana stain

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Challa Sundaram

2014-01-01

Full Text Available Background: Dematiaceous fungi appear brown in tissue section due to melanin in their cell walls. When the brown color is not seen on routine H and E and culture is not available, differentiation of dematiaceous fungi from other fungi is difficult on morphology alone. Aims and Objective: To study if melanin production by dematiaceous fungi can help differentiate them from other types of fungi. Materials and Methods: Fifty tissue sections of various fungal infections and 13 smears from cultures of different species of fungi were stained with Masson Fontana stain to assess melanin production. The tissue sections included biopsies from 26 culture-proven fungi and 24 biopsies of filamentous fungi diagnosed on morphology alone with no culture confirmation. Results: All culture-proven dematiaceous fungi and Zygomycetes showed strong positivity in sections and culture smears. Aspergillus sp showed variable positivity and intensity. Cryptococcus neoformans showed strong positivity in tissue sections and culture smears. Tissue sections of septate filamentous fungi (9/15, Zygomycetes (4/5, and fungi with both hyphal and yeast morphology (4/4 showed positivity for melanin. The septate filamentous fungi negative for melanin were from biopsy samples of fungal sinusitis including both allergic and invasive fungal sinusitis and colonizing fungal balls. Conclusion: Melanin is produced by both dematiaceous and non-dematiaceous fungi. Masson-Fontana stain cannot reliably differentiate dematiaceous fungi from other filamentous fungi like Aspergillus sp; however, absence of melanin in the hyphae may be used to rule out dematiaceous fungi from other filamentous fungi. In the differential diagnosis of yeast fungi, Cryptococcus sp can be differentiated from Candida sp by Masson-Fontana stain in tissue sections.

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

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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

Modified Field's staining--a rapid stain for Trichomonas vaginalis.

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Afzan, M Yusuf; Sivanandam, S; Kumar, G Suresh

2010-10-01

Trichomonas vaginalis, a flagellate protozoan parasite commonly found in the human genitourinary tract, is transmitted primarily by sexual intercourse. Diagnosis is usually by in vitro culture method and staining with Giemsa stain. There are laboratories that use Gram stain as well. We compared the use of modified Field's (MF), Giemsa, and Gram stains on 2 axenic and xenic isolates of T. vaginalis, respectively. Three smears from every sediment of spun cultures of all 4 isolates were stained, respectively, with each of the stains. We showed that MF staining, apart from being a rapid stain (20 s), confers sharper staining contrast, which differentiates the nucleus and the cytoplasm of the organism when compared to Giemsa and Gram staining especially on parasites from spiked urine samples. The alternative staining procedure offers in a diagnostic setting a rapid stain that can easily visualize the parasite with sharp contrasting characteristics between organelles especially the nucleus and cytoplasm. Vacuoles are more clearly visible in parasites stained with MF than when stained with Giemsa. Copyright © 2010 Elsevier Inc. All rights reserved.

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

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)

Gram staining apparatus for space station applications

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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.

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

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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.

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.

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

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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.

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

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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.

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

Jeotgalibacillus soli sp. nov., a Gram-stain-positive bacterium isolated from soil.

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Cunha, Sofia; Tiago, Igor; Paiva, Gabriel; Nobre, Fernanda; da Costa, Milton S; Veríssimo, António

2012-03-01

A Gram-staining-positive, motile, rod-shaped, spore-forming bacterium, designated P9(T), was isolated from soil in Portugal. This organism was aerobic and catalase- and oxidase-positive. It had an optimum growth temperature of about 35 °C and an optimum growth pH of about 8.0-8.5, and grew in medium with 0-9% (w/v) NaCl. The cell-wall peptidoglycan was of the A1α type, with L-lysine as the diagnostic diamino acid. The major respiratory quinone was menaquinone 7 (MK-7) and the major fatty acids were anteiso-C(15:0) (45.4%), iso-C(15:0) (22.0%) and anteiso-C(17:0) (11.2%). The genomic DNA G+C content was about 39.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain P9(T) was most closely related to Jeotgalibacillus campisalis DSM 18983(T) (96.8%) and Jeotgalibacillus marinus DSM 1297(T) (96.5%). These two recognized species formed a coherent cluster with strain P9(T) that was supported by a bootstrap value of 99%. On the basis of the phylogenetic analysis and physiological and biochemical characteristics, strain P9(T) (=DSM 23228(T)=LMG 25523(T)) represents a novel species of the genus Jeotgalibacillus, for which the name Jeotgalibacillus soli sp. nov. is proposed.

Factors influencing extract of Hibiscus sabdariffa staining of rat testes.

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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.

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.

Fungal Fluorescence in Hematoxylin-Eosin Stained Sections

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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

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

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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.

Positive semidefinite matrix completion, universal rigidity and the Strong Arnold Property

NARCIS (Netherlands)

M. Laurent (Monique); A. Varvitsiotis (Antonios)

2014-01-01

htmlabstractThis paper addresses the following three topics: positive semidefinite (psd) matrix completions, universal rigidity of frameworks, and the Strong Arnold Property (SAP). We show some strong connections among these topics, using semidefinite programming as unifying theme. Our main

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.

Positive semidefinite matrix completion, universal rigidity and the Strong Arnold Property

NARCIS (Netherlands)

Laurent, Monique; Varvitsiotis, A.

This paper addresses the following three topics: positive semidefinite (psd) matrix completions, universal rigidity of frameworks, and the Strong Arnold Property (SAP). We show some strong connections among these topics, using semidefinite programming as unifying theme. Our main contribution is a

leaves extracts as counter stain in gram staining reaction 56

African Journals Online (AJOL)

DR. AMINU

is a stain with color contrasting to the principal stain, making the stained ... technology today, the Gram's staining method remains ... was aimed at employing the use of Henna leaves extract as ... fragrant, white or rose flowers in clusters. It is.

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

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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.

[Frequency of intestinal microsporidian infections in HIV-positive patients, as diagnosis by quick hot Gram chromotrope staining and PCR].

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Botero, Jorge H; Montoya, Martha Nelly; Vanegas, Adriana Lucía; Díaz, Abel; Navarro-i-Martínez, Luis; Bornay, Fernando Jorge; Izquierdo, Fernando; del Aguila, Carmen; Agudelo, Sonia del Pilar

2004-12-01

Microsporidia are intracellular obligate parasites, today mainly associated with diarrhea in AIDS patients. Microsporidia prevalence ranges from 8% to 52% in different countries, as evaluated by several diagnostic methods, such as the stain test and PCR. In Medellín, Colombia, its frequency is unknown, and hence, a study was undertaken to determine the frequency of intestinal microsporidiosis in HIV patients, by means of the quick-hot Gram chromotrope test and the PCR. A prospective and descriptive study of an intentional population of all HIV-positive patients was sent to the Grupo Interdisciplinario para el Estudio de las Parasitosis Intestinales laboratory by institutions treating the HIV-positive patients of Medellín between August 2001 and September 2002. The clinical-epidemiological survey included a serial stool test with direct concentration and special stains for coccidiae and intestinal microsporidia. In addition, counts of lymphocytes TCD4+ and viral load were requested. One hundred and three patients with ages ranging from 2-74 years were evaluated. Seventy percent presented with diarrhea--mostly in men (83.5%). The overall frequency of intestinal microsporidiosis was 3.9% and that of other intestinal parasitic infections was 39.8%. Three of the four patients positive for microsporida were infected with Enterocytozoon bieneusi and one with Encephalitozoon intestinalis. The microsporidiosis frequency was relatively low with 3 of the 4 cases associated with protracted diarrhea, counts of LTCD4+ below 100 cel/microl and viral loads up to 100,000 copies.

Port-Wine Stains

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... Safe Videos for Educators Search English Español Port-Wine Stains KidsHealth / For Parents / Port-Wine Stains What's ... Manchas de vino de oporto What Are Port-Wine Stains? A port-wine stain is a type ...

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.

Immunohistochemical staining of precursor forms of prostate-specific antigen (proPSA) in metastatic prostate cancer.

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Parwani, Anil V; Marlow, Cameron; Demarzo, Angelo M; Mikolajczyk, Stephen D; Rittenhouse, Harry G; Veltri, Robert W; Chan, Theresa Y

2006-10-01

Precursors of prostate-specific antigen (proPSA) have been previously shown to be more concentrated in prostate cancer tissue. This study characterizes the immunohistochemical staining (IHS) of proPSA forms in metastatic prostate cancer compared with prostate specific antigen (PSA) and prostatic acid phosphatase (PAP). A tissue microarray, consisting of 74 cases of metastatic prostate carcinoma and control tissues, was used. IHS, using monoclonal antibodies against proPSA with a truncated proleader peptide containing 2 amino acids ([-2]pPSA), native ([-5/-7]pPSA), PSA, and PAP, was analyzed. The monoclonal antibodies were specific for both benign and malignant prostatic glandular tissue. IHS with [-5/-7]pPSA showed the least number of cases with negative staining (3%), and the most number of cases with moderate or strong staining (76%). In the 60 cases where all 4 stains could be evaluated, none of them were negative for proPSA and positive for PSA or PAP, and all 7 cases that were negative for both PSA and PAP showed IHS to proPSA. [-5/-7]pPSA (native proPSA) may be a better marker than PSA and PAP in characterizing metastatic prostate adenocarcinoma, with most of the cases showing positivity for the marker. Even cases that were negative for PSA and PAP, were reactive for proPSA. Such enhanced detection is particularly important in poorly differentiated carcinomas involving metastatic sites where prostate carcinoma is a consideration. A panel of markers, including proPSA, should be performed when metastatic prostate carcinoma is in the differential diagnosis.

Efficacy of in-house fluorescent stain for fungus

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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.

Gram staining in the diagnosis of acute septic arthritis.

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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.

A new technique for Gram staining paraffin-embedded tissue.

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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

Pulmonary infection by Rhodococcus equi presenting with positive Ziehl-Neelsen stain in a patient with human immunodeficiency virus: a case report

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Spiliopoulou, Anastasia; Assimakopoulos, Stelios F; Foka, Antigoni; Kolonitsiou, Fevronia; Lagadinou, Maria; Petinaki, Efthimia; Anastassiou, Evangelos D; Spiliopoulou, Iris; Marangos, Markos

2014-01-01

Introduction Patients with human immunodeficiency virus carry a significant risk of contracting opportunistic infections. The worldwide increased incidence of tuberculosis has instituted pulmonary tuberculosis as an important diagnostic consideration in patients with human immunodeficiency virus presenting with lower respiratory tract infection. A positive result on the readily-available Ziehl-Neelsen stain usually leads to the initiation of antituberculous treatment, since tuberculosis may e...

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

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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.

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

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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.

Evaluation of immunoperoxidase staining technique in the diagnosis of Acanthamoeba keratitis

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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

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

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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

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

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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.

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

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Goto, N

1987-09-01

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

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

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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.

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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.

Metastatic breast cancer to the liver with hepatoid features and Hep Par 1 antibody positive mimicking hepatocellular carcinoma.

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Affleck, Authur; Lyman, William B; Jacobs, W Carl; Livasy, Chad A; Martinie, John B; Iannitti, David A; Vrochides, Dionisios

2018-05-09

The hepatocyte paraffin 1 antibody (Hep Par 1) has a high positive predictive value for differentiating hepatocellular carcinoma from cholangiocarcinoma and metastatic carcinoma. 1 We report a case of metastatic breast cancer to the liver with hepatoid histology and strong positive staining for Hep Par 1 mimicking hepatocellular carcinoma. To our knowledge, primary breast carcinoma staining Hep Par 1 positive has not been reported in the setting of hepatic metastasis. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

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

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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.

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

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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.

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.

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

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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.

A case of perforated sigmoid diverticulitis in which gram staining of ascitic fluid was useful for diagnosis.

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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.

[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.

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

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

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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.

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

Facile method to stain the bacterial cell surface for super-resolution fluorescence microscopy

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Gunsolus, Ian L.; Hu, Dehong; Mihai, Cosmin; Lohse, Samuel E.; Lee, Chang-Soo; Torelli, Marco; Hamers, Robert J.; Murphy, Catherine; Orr, Galya; Haynes, Christy L.

2014-01-01

A method to fluorescently stain the surfaces of both Gram-negative and Gram-positive bacterial cells compatible with super-resolution fluorescence microscopy is presented. This method utilizes a commercially-available fluorescent probe to label primary amines at the surface of the cell. We demonstrate efficient staining of two bacterial strains, the Gram-negative Shewanella oneidensis MR-1 and the Gram-positive Bacillus subtilis 168. Using structured illumination microscopy and stochastic optical reconstruction microscopy, which require high quantum yield or specialized dyes, we show that this staining method may be used to resolve the bacterial cell surface with sub-diffraction-limited resolution. We further use this method to identify localization patterns of nanomaterials, specifically cadmium selenide quantum dots, following interaction with bacterial cells.

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

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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.

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

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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.

Metabolic markers in relation to hypoxia; staining patterns and colocalization of pimonidazole, HIF-1α, CAIX, LDH-5, GLUT-1, MCT1 and MCT4

International Nuclear Information System (INIS)

Rademakers, Saskia E; Lok, Jasper; Kogel, Albert J van der; Bussink, Johan; Kaanders, Johannes HAM

2011-01-01

The cellular response of malignant tumors to hypoxia is diverse. Several important endogenous metabolic markers are upregulated under hypoxic conditions. We examined the staining patterns and co-expression of HIF-1α, CAIX, LDH-5, GLUT-1, MCT1 and MCT4 with the exogenous hypoxic cell marker pimonidazole and the association of marker expression with clinicopathological characteristics. 20 biopsies of advanced head and neck carcinomas were immunohistochemically stained and analyzed. All patients were given the hypoxia marker pimonidazole intravenously 2 h prior to biopsy taking. The tumor area positive for each marker, the colocalization of the different markers and the distribution of the markers in relation to the blood vessels were assessed by semiautomatic quantitative analysis. MCT1 staining was present in hypoxic (pimonidazole stained) as well as non-hypoxic areas in almost equal amounts. MCT1 expression showed a significant overall correlation (r = 0.75, p < 0.001) and strong spatial relationship with CAIX. LDH-5 showed the strongest correlation with pimonidazole (r = 0.66, p = 0.002). MCT4 and GLUT-1 demonstrated a typical diffusion-limited hypoxic pattern and showed a high degree of colocalization. Both MCT4 and CAIX showed a higher expression in the primary tumor in node positive patients (p = 0.09 both). Colocalization and staining patterns of metabolic and hypoxia-related proteins provides valuable additional information over single protein analyses and can improve the understanding of their functions and environmental influences

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...

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

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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.

Alternate gram staining technique using a fluorescent lectin.

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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

Impact of round-the-clock CSF Gram stain on empirical therapy for suspected central nervous system infections.

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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.

Position-Dependent Dynamics Explain Pore-Averaged Diffusion in Strongly Attractive Adsorptive Systems.

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Krekelberg, William P; Siderius, Daniel W; Shen, Vincent K; Truskett, Thomas M; Errington, Jeffrey R

2017-12-12

Using molecular simulations, we investigate the relationship between the pore-averaged and position-dependent self-diffusivity of a fluid adsorbed in a strongly attractive pore as a function of loading. Previous work (Krekelberg, W. P.; Siderius, D. W.; Shen, V. K.; Truskett, T. M.; Errington, J. R. Connection between thermodynamics and dynamics of simple fluids in highly attractive pores. Langmuir 2013, 29, 14527-14535, doi: 10.1021/la4037327) established that pore-averaged self-diffusivity in the multilayer adsorption regime, where the fluid exhibits a dense film at the pore surface and a lower density interior pore region, is nearly constant as a function of loading. Here we show that this puzzling behavior can be understood in terms of how loading affects the fraction of particles that reside in the film and interior pore regions as well as their distinct dynamics. Specifically, the insensitivity of pore-averaged diffusivity to loading arises from the approximate cancellation of two factors: an increase in the fraction of particles in the higher diffusivity interior pore region with loading and a corresponding decrease in the particle diffusivity in that region. We also find that the position-dependent self-diffusivities scale with the position-dependent density. We present a model for predicting the pore-average self-diffusivity based on the position-dependent self-diffusivity, which captures the unusual characteristics of pore-averaged self-diffusivity in strongly attractive pores over several orders of magnitude.

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

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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.

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.

Determination of ABO blood grouping from human oral squamous epithelium by the highly sensitive immunohistochemical staining method EnVision+.

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Noda, Hiroshi; Yokota, Makoto; Tatsumi, Shinji; Sugiyama, Shizuyuki

2002-03-01

Using the highly sensitive immunohistochemical staining method EnVision+, which employs a dextran polymer reagent for the secondary antibody, the detection of the ABH antigens was attempted in the oral squamous epithelium. This new technique uses monoclonal antibody as a primary antibody and it takes about three hours for staining. The time is much shorter than conventional absorption-elution testing or absorption-inhibition testing for the determination of ABO blood grouping. Secretor saliva samples were stained at strong intensity by the antibody, which corresponded to its blood group and anti-H. On the one hand, nonsecretor saliva samples were stained at strong intensity only by the antibody that corresponded to its blood group, and at weak intensity only by anti-H. Since human oral squamous epithelium antigens were stained specifically by this method, we can examine the ABO blood group of saliva samples and perform cytodiagnosis at the same time. Our research suggested that the EnVision+ Method is a useful technique for ABO blood grouping of saliva in forensic cases.

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

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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.

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

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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.

Automated Interpretation of Blood Culture Gram Stains by Use of a Deep Convolutional Neural Network.

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Smith, Kenneth P; Kang, Anthony D; Kirby, James E

2018-03-01

Microscopic interpretation of stained smears is one of the most operator-dependent and time-intensive activities in the clinical microbiology laboratory. Here, we investigated application of an automated image acquisition and convolutional neural network (CNN)-based approach for automated Gram stain classification. Using an automated microscopy platform, uncoverslipped slides were scanned with a 40× dry objective, generating images of sufficient resolution for interpretation. We collected 25,488 images from positive blood culture Gram stains prepared during routine clinical workup. These images were used to generate 100,213 crops containing Gram-positive cocci in clusters, Gram-positive cocci in chains/pairs, Gram-negative rods, or background (no cells). These categories were targeted for proof-of-concept development as they are associated with the majority of bloodstream infections. Our CNN model achieved a classification accuracy of 94.9% on a test set of image crops. Receiver operating characteristic (ROC) curve analysis indicated a robust ability to differentiate between categories with an area under the curve of >0.98 for each. After training and validation, we applied the classification algorithm to new images collected from 189 whole slides without human intervention. Sensitivity and specificity were 98.4% and 75.0% for Gram-positive cocci in chains and pairs, 93.2% and 97.2% for Gram-positive cocci in clusters, and 96.3% and 98.1% for Gram-negative rods. Taken together, our data support a proof of concept for a fully automated classification methodology for blood-culture Gram stains. Importantly, the algorithm was highly adept at identifying image crops with organisms and could be used to present prescreened, classified crops to technologists to accelerate smear review. This concept could potentially be extended to all Gram stain interpretive activities in the clinical laboratory. Copyright © 2018 American Society for Microbiology.

Staining for factor VIII related antigen and Ulex europaeus agglutinin I (UEA-I) in 230 tumours. An assessment of their specificity for angiosarcoma and Kaposi's sarcoma.

Science.gov (United States)

Leader, M; Collins, M; Patel, J; Henry, K

1986-11-01

In this study we examined the staining reactivity of commercially available antisera to factor VIII related antigen (F VIII RAg) and Ulex europaeus agglutinin I (UEA-I) on sections from 230 formalin fixed paraffin embedded tumours. These included 196 sarcomas, 20 carcinomas and 14 angiomas. All angiomas showed positive staining for F VIII RAg; all carcinomas showed negative staining; the vasoformative areas of all angiosarcomas stained positively but only four of six angiosarcomas showed positive staining of their solid areas; of seven Kaposi's sarcomas, all showed positive staining of vessels and six showed positive staining of the spindle cell component. In the remaining 181 non-vascular sarcomas there was a false positive result in four tumours (2.2%), three of which had a history of irradiation. Pre-radiotherapy biopsies of these three tumours stained negatively with anti-F VIII RAg. UEA-I was demonstrated in all the angiomas studied, in all angiosarcomas (including the solid components) and in well-formed vessels of all Kaposi's sarcomas, but only in the spindle cell component of 3/6. However, there was an unacceptably high rate of false positive staining amongst the carcinomas and non-vascular sarcomas. In conclusion, F VIII RAg is a specific but not a sensitive marker of angiosarcomas; UEA-I is a sensitive but not a specific marker of angiosarcomas.

Utility of Iron Staining in Identifying the Cause of Renal Allograft Dysfunction in Patients with Sickle Cell Disease

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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.

Simultaneous fluorescent gram staining and activity assessment of activated sludge bacteria.

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Forster, Scott; Snape, Jason R; Lappin-Scott, Hilary M; Porter, Jonathan

2002-10-01

Wastewater treatment is one of the most important commercial biotechnological processes, and yet the component bacterial populations and their associated metabolic activities are poorly understood. The novel fluorescent dye hexidium iodide allows assessment of Gram status by differential absorption through bacterial cell walls. Differentiation between gram-positive and gram-negative wastewater bacteria was achieved after flow cytometric analysis. This study shows that the relative proportions of gram-positive and gram-negative bacterial cells identified by traditional microscopy and hexidium iodide staining were not significantly different. Dual staining of cells for Gram status and activity proved effective in analyzing mixtures of cultured bacteria and wastewater populations. Levels of highly active organisms at two wastewater treatment plants, both gram positive and gram negative, ranged from 1.5% in activated sludge flocs to 16% in the activated sludge fluid. Gram-positive organisms comprised Gram status and activity within activated sludge samples over a 4-day period showed significant differences over time. This method provides a rapid, quantitative measure of Gram status linked with in situ activity within wastewater systems.

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

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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.

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.

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

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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.

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

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Netuschil, Lutz; Auschill, Thorsten M; Sculean, Anton; Arweiler, Nicole B

2014-01-11

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

Compared With Elastin Stains, h-Caldesmon and Desmin Offer Superior Detection of Vessel Invasion in Gastric, Pancreatic, and Colorectal Adenocarcinomas.

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Ekinci, Özgür; Öğüt, Betül; Çelik, Bülent; Dursun, Ayşe

2018-06-01

The presence of vessel invasion is considered indicative of a poor prognosis in many malignant tumors. We aimed to compare the sensitivity of elastin stains (van Gieson's and orcein methods) with 2 smooth muscle markers (h-caldesmon and desmin) in gastric, pancreatic, and colorectal adenocarcinoma specimens. We used 27 (29.3%) gastric, 35 (38.0%) pancreatic, and 30 (32.6%) colorectal resection specimens. We applied a provisional classification of vessel invasion patterns: type A, a focus with a nearby artery unaccompanied by a vein; type T, a focus at the invasive front without an unaccompanied artery; and type X, foci that only appeared by any of the 4 stains used. There were 369 foci. The smooth muscle markers were more sensitive than the elastin stains, and h-caldesmon more sensitive than desmin, in all types. Among the 139 type A foci, 33 (23.7%) were positive by desmin and h-caldesmon, whereas the elastin stains were not ( P = .001). h-Caldesmon was the only positive marker in 11 (7.9%; P = .011). Among the 78 type T foci, 21 (26.9%) were positive by desmin and h-caldesmon, when both elastin stains were negative ( P = .000). In 16 (20.5%) foci, h-caldesmon was the only positive marker ( P = .002). Among 152 type X foci, 91 (59.9%) were positive by all markers, 26 (17.1%) by both desmin and h-caldesmon, and 9 (5.9%) by only the 2 elastin stains ( P = .001). We recommend these stains for suspect foci in gastric, pancreatic, and colorectal adenocarcinoma specimens. They might highlight both predictable and unpredictable foci.

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

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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.

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

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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.

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.

c-KIT positive schistosomal urinary bladder carcinoma are frequent but lack KIT gene mutations.

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Shams, Tahany M; Metawea, Mokhtar; Salim, Elsayed I

2013-01-01

Urinary bladder squamous cell carcinoma (SCC), one of the most common neoplasms in Egypt, is attributed to chronic urinary infection with Schistosoma haematobium (Schistosomiasis). The proto-oncogene c-KIT, encoding a tyrosine kinase receptor and implicated in the development of a number of human malignancies, has not been studied so far in schistosomal urinary bladder SCCs. We therefore determined immunohistochemical (IHC) expression of c-KIT in paraffin sections from 120 radical cystectomies of SCCs originally obtained from the Pathology Department of Suez Canal University (Ismailia, Egypt). Each slide was evaluated for staining intensity where the staining extent of >10% of cells was considered positive. c-KIT overexpression was detected in 78.3% (94/120) of the patients, the staining extents in the tumor cells were 11-50% and >50% in 40 (42.6%) and 54 (57.4%) respectively. The positive cases had 14.9%, 63.8%, 21.3% as weak, moderate and strong intensity respectively. Patients with positive bilharzial ova had significantly higher c-KIT expression than patients without (95.2% vs. 38.9%, P=0.000). Mutation analysis of exons 9-13 was negative in thirty KIT positive cases. The high rate of positivity in SBSCC was one of the striking findings; However, CD117 may be a potential target for site specific immunotherapy to improve the outcome of this tumor.

Routine Hematoxylin and Eosin Stain Is Specific for the Diagnosis of Cytomegalovirus Infection in Gastrointestinal Biopsy Specimens.

Science.gov (United States)

Guo, Ling; DeRoche, Tom C; Salih, Ziyan T; Qasem, Shadi A

2018-03-01

Gastrointestinal cytomegalovirus (CMV) infection is a serious complication in immunocompromised patients; clinicians often expect expedited results for biopsy specimens. Our goal is to determine the accuracy of identification of CMV on hematoxylin and eosin (H&E) stain. A total of 361 biopsy specimens from 273 patients with suspicion for CMV infection were retrieved. CMV was detected by immunohistochemistry (IHC) in 37 specimens acquired from 33 individual patients (average age = 54 years). Among the CMV-positive patients, 29 (88%) were reported to be immunosuppressed. Colon was the most common affected location. Of 37 CMV-positive specimens by IHC, 28 were positive by H&E (76%), 6 were negative (16%), and 3 were suspicious (8%). Of the 29 positive specimens on H&E, 28 were confirmed by IHC (97%) and 1 was indeterminate (3%). The sensitivity and specificity of H&E were 84% and 94%, respectively; the positive predictive value was 97%, and the negative predictive value was 93% ( P < .00001). Our results show that a preliminary diagnosis of CMV infection, based on H&E stains, can be reported with high specificity and low risk for false-positive results. Suspicious cases should be deferred pending the result of IHC stains.

Iron Stain on Wood

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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...

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.

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.

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.

The effect of corrosion on stained glass windows

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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

Centrifuge-operated specimen staining method and apparatus

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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.

[Application of Immunohistochemistry and Immunofluorescence Staining in Detection of Phospholipase A2 Receptor on Paraffin Section of Renal Biopsy Tissue].

Science.gov (United States)

Dong, Hong-rui; Wang, Yan-yan; Wang, Guo-qin; Sun, Li-jun; Cheng, Hong; Chen, Yi-pu

2015-10-01

To evaluate the application of immunohistochemistry and fluorescence staining method in the detection of phospholipase A2 receptor (PLA2R) on paraffin section of renal biopsy tissue,and to find an accurate and fast method for the detection of PLA2R in renal tissue. The PLA2R of 193 cases were detected by immunohistochemical staining,and the antigen was repaired by the method of high pressure cooker (HPC) hot repair plus trypsin repair. The 193 samples including 139 cases of idiopathic membranous nephropathy (IMN), 15 cases of membranous lupus nephritis, 8 cases of hepatitis B virus associated membranous nephropathy, 18 cases of IgA nephropathy, and 13 cases of minimal change diseases. To compare the dyeing effects, 22 paraffin sections of renal biopsy tissue of IMN cases with positive PLA2R were stained by using 4 different. of antigen repairing,which included HPC hot repair, HPC hot repair plus trypsin repair, water bath heat repair, and water bath heat repair plus trypsin repair. To compare the dyeing effects, 15 paraffin sections of renal biopsy tissue of IMN cases with positive PLA2R were stained by using 3 different. of antigen repairing,which included water bath heat repair plus trypsin repair, protease K digestion repair, and pepsin digestion repair. In 193 cases, the positive rate of PLA2R in IMN cases was 90.6% (126/139), and the other 54 patients without IMN were negative. Twenty-two IMN patients were positive for PLA2R by using the HPC heat repair plus trypsin repaire or the water bath heat repair plus trypsin repair;while only a few cases of 22 IMN cases were positive by using the HPC hot repair alone or water bath heat repair alone. Fifteen IMN patients were positive for PLA2R by using water bath heat repair plus trypsin repair,protease K digestion repair,and pepsin digestion repair, but the distribution of positive deposits and the background were different. PLA2R immunohistochemical staining can effectively identify IMN and secondary MN. For

The accuracy of Gram stain of respiratory specimens in excluding Staphylococcus aureus in ventilator-associated pneumonia.

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Gottesman, Tamar; Yossepowitch, Orit; Lerner, Evgenia; Schwartz-Harari, Orna; Soroksky, Arie; Yekutieli, Daniel; Dan, Michael

2014-10-01

To evaluate the Gram stain of deep tracheal aspirate as a tool to direct empiric antibiotic therapy, and more specifically as a tool to exclude the need for empiric antibiotic coverage against Staphylococcus aureus in ventilator-associated pneumonia (VAP). A prospective, single-center, observational, cohort study. All wards at a community hospital. Adult patients requiring mechanical ventilation, identified as having VAP in a 54-month prospective surveillance database. Sampling of lower airway secretions by deep endotracheal aspiration was taken from each patient who developed VAP. Samples were sent immediately for Gram stain and qualitative bacterial cultures. Demographic and relevant clinical data were collected; Gram stain, culture, and antibiotic susceptibility results were documented; and outcome was followed prospectively. The analysis included 114 consecutive patients with 115 episodes of VAP from June 2007 to January 2012. Sensitivity of Gram stain compared with culture was 90.47% for gram-positive cocci, 69.6% for gram-negative rods, and 50% for sterile cultures. Specificity was 82.5%, 77.8%, and 79%, respectively. Negative predictive value was high for gram-positive cocci (97%) and sterile cultures (96%) but low for gram-negative rods (20%). Acinetobacter baumanii (45%) and Pseudomonas aeruginosa (38 %) were the prevailing isolates. S aureus was found in 18.3% of the patients. Most isolates were multiresistant. Absence of gram-positive bacteria on Gram stain had a high negative predictive value. These data can be used to narrow the initial empiric antibiotic regimen and to avoid unnecessary exposure of patients to vancomycin and other antistaphyloccocal agents. Copyright © 2014 Elsevier Inc. All rights reserved.

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

DEFF Research Database (Denmark)

Prentø, P

1993-01-01

this and experiments with additives (sodium dodecylsulphate, urea etc.) and organic solvents, it is proposed that coagulant interchain cross-linking at the high protein concentration of the cytoplasm masks potential dye-binding sites. This affects high affinity dyes with multiple binding sites more than small dyes......, and so puts AcF at a disadvantage compared to PA. Staining of non-collagen proteins is mainly by hydrophobic bonding, involving ionic attractions, apolar bonds, and release of water. This mode of binding is relatively strong, decreases swelling and leads to slow dye exchange. Dye binding to collagen...

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.

Secretory leukocyte protease inhibitor expression and high-risk HPV infection in anal lesions of HIV positive patients

Science.gov (United States)

NUOVO, Gerard J.; GRINSZTEJN, Beatriz; FRIEDMAN, Ruth K.; VELOSO, Valdiléa G.; CUNHA, Cynthia B.; COUTINHO, José R.; VIANNA-ANDRADE, Cecilia; OLIVEIRA, Nathalia S.; WOODHAM, Andrew W.; DA SILVA, Diane M.; KAST, W. Martin

2016-01-01

Objective The aim of the current study was to evaluate secretory leukocyte protease inhibitor (SLPI) expression in anal biopsies from HIV-positive (HIV+) individuals, and compare that to anal intraepithelial neoplasia (AIN) diagnoses and human papillomavirus (HPV) status. Design This is a cross-sectional study of a cohort of 54 HIV+ (31 males and 23 females) from an AIDS clinic in Rio de Janeiro, Brazil. Methods The study material consisted of anorectal tissue biopsies obtained from HIV+ subjects, which were used to construct tissue microarray paraffin blocks for immunohistochemical analysis of SLPI expression. Biopsies were evaluated by an expert pathologist and classified as low-grade anal intraepithelial neoplasia (AIN1), high-grade anal intraepithelial neoplasia (AIN2/3), or normal squamous epithelium. Additionally, DNA from the biopsies was extracted and analyzed for the presence of low- or high-risk HPV DNA. Results Histologically normal squamous epithelium from the anorectal region showed strong positive SLPI staining in 17/20 (85%) samples. In comparison, 9/17 (53%) dysplastic squamous epithelial samples from AIN1 patients showed strong SLPI staining, and only 5/17 (29%) samples from AIN2-3 patients exhibited strong SPLI staining, which both were significantly fewer than those from normal tissue (p=0.005). Furthermore, there was a significantly higher proportion of samples in which oncogenic high-risk HPV genotypes were detected in low SLPI expressing tissues than that in tissues with high SLPI expression (p=0.040). Conclusion Taken together these results suggest that low SLPI expression is associated with high-risk HPV infections in the development of AIN. PMID:27149102

Comparison of three staining methods for the detection of intestinal microspora spp.

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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.

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

Study of bacterial meningitis in children below 5 years with comparative evaluation of gram staining, culture and bacterial antigen detection.

Science.gov (United States)

Yadhav Ml, Kala

2014-04-01

Bacterial meningitis is one of the most serious infections seen in infants and children, which is associated with acute complications and chronic morbidity. Infections of Central Nervous System (CNS) still dominate the scene of childhood neurological disorders in most of the developing tropical countries. To isolate, identify and determine the antibiotic susceptibility patterns of pathogens associated with bacterial meningitis. We also aimed to comparatively evaluate of Gram staining, culture and bacterial antigen detection in cerebrospinal fluid samples. Present comparative study included 100 CSF samples of children below the age of 5 years, who were clinically suspected meningitis cases. The samples were subjected to Gram staining, culture and Latex agglutination test (LAT). The organisms isolated in the study were characterized and antibiotic susceptibility test was done according to standard guidelines. It was done by using Gaussian test. Of the 100 cases, 24 were diagnosed as Acute bacterial meningitis (ABM) cases by. Gram staining, culture and latex agglutination test. 21 (87.5%) cases were culture positive, with 2 cases being positive for polymicrobial isolates. Gram staining was positive in 17 (70.53%) cases and LAT was positive in 18 (33.33%) cases. Streptococcus pneumoniae was the predominant organism which was isolated and it was sensitive to antibiotics. In the present study, male to female ratio was 1.27:1, which showed a male preponderance. With the combination of Gram staining, culture, and LAT, 100% sensitivity and specificity can be achieved (p Gram staining and LAT can detect 85% of cases of ABM. Bacterial meningitis is a medical emergency and making an early diagnosis and providing treatment early are life saving and they reduce chronic morbidity.

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

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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.

Pitfalls using tyramide signal amplification (TSA) in the mouse gastrointestinal tract: endogenous streptavidin-binding sites lead to false positive staining.

Science.gov (United States)

Horling, L; Neuhuber, W L; Raab, M

2012-02-15

Highly sensitive immunohistochemical detection systems such as tyramide signal amplification (TSA) are widely used, since they allow using two primary antibodies raised in the same species. Most of them are based on the streptavidin-biotin-peroxidase system and include streptavidin-coupled secondary antibodies. Using TSA in cryostat-sectioned tissues of mouse esophagus, we were puzzled by negative controls with unexpected staining mostly in the ganglionic areas. This prompted us to search for the causing agent and to include also other parts of the mouse gastrointestinal tract for comparison. Streptavidin-coupled antibodies bound to endogenous binding sites yet to be characterized, which are present throughout the mouse intestines. Staining was mainly localized around neuronal cell bodies of enteric ganglia. Thus, caution is warranted when applying streptavidin-coupled antibodies in the mouse gastrointestinal tract. The use of endogenous biotin-blocking kits combined with a prolonged post-fixation time could significantly reduce unintentional staining. Copyright © 2011 Elsevier B.V. All rights reserved.

Histopathological evaluation of ocular microsporidiosis by different stains

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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.

An automated cell-counting algorithm for fluorescently-stained cells in migration assays

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Novielli Nicole M

2011-10-01

Full Text Available Abstract A cell-counting algorithm, developed in Matlab®, was created to efficiently count migrated fluorescently-stained cells on membranes from migration assays. At each concentration of cells used (10,000, and 100,000 cells, images were acquired at 2.5 ×, 5 ×, and 10 × objective magnifications. Automated cell counts strongly correlated to manual counts (r2 = 0.99, P

Visual evoked potentials show strong positive association with intracranial pressure in patients with cryptococcal meningitis

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Marcelo Adriano da Cunha Silva Vieira

2015-04-01

Full Text Available Objective : To verify the relationship between intracranial pressure and flash visual evoked potentials (F-VEP in patients with cryptococcal meningitis. Method The sample included adults diagnosed with cryptococcal meningitis admitted at a reference hospital for infectious diseases. The patients were subjected to F-VEP tests shortly before lumbar puncture. The Pearson’s linear correlation coefficient was calculated and the linear regression analysis was performed. Results : Eighteen individuals were subjected to a total of 69 lumbar punctures preceded by F-VEP tests. At the first lumbar puncture performed in each patient, N2 latency exhibited a strong positive correlation with intracranial pressure (r = 0.83; CI = 0.60 - 0.94; p < 0.0001. The direction of this relationship was maintained in subsequent punctures. Conclusion : The intracranial pressure measured by spinal tap manometry showed strong positive association with the N2 latency F-VEP in patients with cryptococcal meningitis.

The use of Gram stain and matrix-assisted laser desorption ionization time-of-flight mass spectrometry on positive blood culture: synergy between new and old technology.

Science.gov (United States)

Fuglsang-Damgaard, David; Nielsen, Camilla Houlberg; Mandrup, Elisabeth; Fuursted, Kurt

2011-10-01

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is promising as an alternative to more costly and cumbersome methods for direct identifications in blood cultures. We wanted to evaluate a simplified pre-treatment method for using MALDI-TOF-MS directly on positive blood cultures using BacT/Alert blood culture system, and to test an algorithm combining the result of the initial microscopy with the result suggested by MALDI-TOF-MS. Using the recommended cut-off score of 1.7 the best results were obtained among Gram-negative rods with correct identifications in 91% of Enterobacteriaceae, 83% in aerobic/non-fermentative Gram-negative rods, whereas results were more modest among Gram-positive cocci with correct identifications in 52% of Staphylococci, 54% in Enterococci and only 20% in Streptococci. Combining the results of Gram stain with the top reports by MALDI-TOF-MS, increased the sensitivity from 91% to 93% in the score range from 1.5 to 1.7 and from 48% to 85% in the score range from 1.3 to 1.5. Thus, using this strategy and accepting a cut-off at 1.3 instead of the suggested 1.7, overall sensitivity could be increased from 88.1% to 96.3%. MALDI-TOF-MS is an efficient method for direct routine identification of bacterial isolates in blood culture, especially when combined with the result of the Gram stain. © 2011 The Authors. APMIS © 2011 APMIS.

Entamoeba spp. diagnosis in patients with inflmmatory diarrhea by staining, copro-antigen ELISA and multiplex PCR methods

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Zahra Gharibi

2017-10-01

Full Text Available Objective: To evaluate Entamoeba spp. diagnosis in patients with inflammatory diarrhea by staining, copro-antigen ELISA and multiplex PCR methods. Methods: In this descriptive cross-sectional survey, 200 stool samples were randomly collected during 2015–2016. The stool samples were evaluated microscopically for the presence of the parasite using direct and formalin-ether concentration and trichrome staining methods. Then, the stool samples were examined by copro-antigen ELISA (Biomerica Company and multiplex PCR methods. Results: Of 200 samples, 17, 29 and 23 cases were positive for Entamoeba species by the staining, copro-antigen ELISA and multiplex PCR methods, respectively. Of 23 positive samples in multiplex PCR test, 13 and 10 samples were positive for Entamoeba dispar (E. dispar and Entamoeba histolytica (E. histolytica, respectively. Conclusions: Our finding indicated a relatively high prevalence of Entamoeba species in patients with inflammatory diarrhea in Ahvaz city. Due to the complications of E. histolytica/ dispar infection, the health authorities of the city must pay more attention to control and prevent the transmission of E. histolytica/dispar to individuals.

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)

Basal cell carcinoma: CD56 and cytokeratin 5/6 staining patterns in the differential diagnosis with Merkel cell carcinoma.

Science.gov (United States)

Panse, Gauri; McNiff, Jennifer M; Ko, Christine J

2017-06-01

Basal cell carcinoma (BCC) can resemble Merkel cell carcinoma (MCC) on histopathological examination and while CK20 is a useful marker in this differential, it is occasionally negative in MCC. CD56, a sensitive marker of neuroendocrine differentiation, is sometimes used to identify MCC, but has been reportedly variably positive in BCC as well. In contrast, CK5/6 consistently labels BCC but is not expressed in neuroendocrine tumors. We evaluated 20 cases of BCC for the pattern of CD56 and cytokeratin 5/6 (CK5/6) staining, hypothesizing that these 2 stains could differentiate BCC from MCC in difficult cases. Seventeen cases of MCC previously stained with CD56 were also examined. All BCCs showed patchy expression of CD56 except for 2 cases, which showed staining of greater than 70% of tumor. CK5/6 was diffusely positive in all cases of BCC. Fifteen of 17 MCCs were diffusely positive for CD56. The difference in the pattern of CD56 expression between MCC and BCC (diffuse vs patchy, respectively) was statistically significant (P < .05). BCC typically shows patchy CD56 expression and diffuse CK5/6 positivity. These 2 markers can be used to distinguish between BCC and MCC in challenging cases. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

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...

Proliferation assessment in breast carcinomas using digital image analysis based on virtual Ki67/cytokeratin double staining.

Science.gov (United States)

Røge, Rasmus; Riber-Hansen, Rikke; Nielsen, Søren; Vyberg, Mogens

2016-07-01

Manual estimation of Ki67 Proliferation Index (PI) in breast carcinoma classification is labor intensive and prone to intra- and interobserver variation. Standard Digital Image Analysis (DIA) has limitations due to issues with tumor cell identification. Recently, a computer algorithm, DIA based on Virtual Double Staining (VDS), segmenting Ki67-positive and -negative tumor cells using digitally fused parallel cytokeratin (CK) and Ki67-stained slides has been introduced. In this study, we compare VDS with manual stereological counting of Ki67-positive and -negative cells and examine the impact of the physical distance of the parallel slides on the alignment of slides. TMAs, containing 140 cores of consecutively obtained breast carcinomas, were stained for CK and Ki67 using optimized staining protocols. By means of stereological principles, Ki67-positive and -negative cell profiles were counted in sampled areas and used for the estimation of PIs of the whole tissue core. The VDS principle was applied to both the same sampled areas and the whole tissue core. Additionally, five neighboring slides were stained for CK in order to examine the alignment algorithm. Correlation between manual counting and VDS in both sampled areas and whole core was almost perfect (correlation coefficients above 0.97). Bland-Altman plots did not reveal any skewness in any data ranges. There was a good agreement in alignment (>85 %) in neighboring slides, whereas agreement decreased in non-neighboring slides. VDS gave similar results compared with manual counting using stereological principles. Introduction of this method in clinical and research practice may improve accuracy and reproducibility of Ki67 PI.

Intraoperative immunohistochemistry staining of sentinel nodes in breast cancer: Clinical and economical implications

DEFF Research Database (Denmark)

Holm, M.; Paaschburg, B.; Balslev, E.

2008-01-01

.0001) for isolated tumor cell metastasis, from 56 to 36.4% (p analysis showed an overall...... of the surgical procedures before (n = 706) and after (n = 503) introducing intraoperatice IHC on frozen section. We also did a cost analysis. Intraoperative IHC staining led to a lowering of the late positive SNB rate. Introducing IHC gave a decrease in the late positive rate from 93 to 52% (p

Utility of Modified Ultrafast Papanicolaou Stain in Cytological Diagnosis.

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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.

Diagnosis of common bacterial causes of urethritis in men by Gram stain, culture and multiplex PCR.

Science.gov (United States)

Jahan, F; Shamsuzzaman, S M; Akter, S

2014-12-01

Urethritis is one of the most important causes of morbidity and mortality in developing countries. The aim of this study was to detect common bacterial causes of urethritis in men by Gram stain, culture and multiplex PCR.185 male patients who presented at the Skin and venereal clinic of the Dhaka Medical College, Bangladesh with clinical symptoms suggestive of urethritis were enrolled in this study. Urethral discharges were tested for detection of Neisseria gonorrhoeae by Gram stain, culture and PCR. Multiplex PCR assay was done to detect DNA of Chlamydia trachomatis, Ureaplasma urealyticum and Mycoplasma genitalium. Out of 185 participants, 30.27% and 14.6% were infected by Neisseria gonorrhoeae and Chlamydia trachomatis respectively. None of the individuals was found positive for either Ureaplasma urealyticum or Mycoplasma genitalium. Among the Neisseria gonorrhoeae positive patients 27.57% were positive from Gram stain, 26.49% were culture positive, 30.27% were positive by PCR (p<0.001). 32.65% of the Neisseria gonorrhoeae isolates were penicillinase producers and 83.67% were susceptible to ceftriaxone. Considering culture as the gold standard, the sensitivity and specificity of PCR for the detection of Neisseria gonorrhoeae was 100%, and 94.85% respectively with an accuracy of 96.22%. 3.73% of the 134 smear negative and 5.15% of the 136 culture negative samples were positive by PCR. PCR was the most sensitive and rapid method for the diagnosis of urethritis. Multiplex PCR may be a useful approach to laboratory diagnosis of urethritis in men for its high sensitivity and specificity.

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.

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

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

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.

Case of Mycobacterium tuberculosis meningitis: Gram staining as a useful initial diagnostic clue for tuberculous meningitis.

Science.gov (United States)

Kawakami, Sayoko; Kawamura, Yasuyosi; Nishiyama, Kyouhei; Hatanaka, Hiroki; Fujisaki, Ryuichi; Ono, Yasuo; Miyazawa, Yukihisa; Nishiya, Hajime

2012-12-01

A 32-year-old man was admitted to our hospital because of fever, headache, and loss of consciousness. Four days before admission, he had had difficulty speaking. On the day of admission, his colleague had found him to be unconscious and lying on his back. He was admitted to our hospital. The temperature at the eardrum was 35.2°C. Neurologic evaluation was negative. Computed tomography (CT) scan of the brain showed slight ventricular enlargement bilaterally. An X-ray film of the chest showed no abnormality. On the second hospital day, neck stiffness was noted. The cerebrospinal fluid (CSF) contained 870 white cells/μl, most of which were neutrophils; the glucose level in the CSF was 10 mg/dl, and the protein level was 140 mg/dl. Stained smears of the CSF, including Gram staining and India-ink preparations, disclosed no microorganisms. Capsular antigen tests for several bacteria were negative. Antimicrobial agents were started. However, by changing the microscope focus slightly while viewing Gram stains of the CSF, we could see brightened and Gram-positive bacilli that had been phagocytosed by neutrophils. This finding suggested the presence of Mycobacterium tuberculosis. Ziehl-Neelsen staining of the CSF and gastric juice revealed anti-acid bacilli. Polymerase chain reaction for M. tuberculosis in the gastric juice was positive. This case showed that Gram staining could be useful as an initial adjunct for the diagnosis of tuberculous meningitis, particularly when the CSF shows predominantly neutrocytic pleocytosis, but no other evidence of bacterial meningitis.

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.

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.

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 ...

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.

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

Use of magnetic beads for Gram staining of bacteria in aqueous suspension.

Science.gov (United States)

Yazdankhah, S P; Sørum, H; Larsen, H J; Gogstad, G

2001-12-01

A Gram staining technique was developed using monodisperse magnetic beads in concentrating bacteria in suspension for downstream application. The technique does not require heat fixation of organisms, electrical power, or a microscope. Gram-negative and Gram-positive bacteria were identified macroscopically based on the colour of the suspension. The bacteria concentrated on magnetic beads may also be identified microscopically.

Self-assembly of highly fluorescent semiconductor nanorods into large scale smectic liquid crystal structures by coffee stain evaporation dynamics

International Nuclear Information System (INIS)

Nobile, Concetta; Carbone, Luigi; Fiore, Angela; Cingolani, Roberto; Manna, Liberato; Krahne, Roman

2009-01-01

We deposit droplets of nanorods dispersed in solvents on substrate surfaces and let the solvent evaporate. We find that strong contact line pinning leads to dense nanorod deposition inside coffee stain fringes, where we observe large scale lateral ordering of the nanorods with the long axis of the rods oriented parallel to the contact line. We observe birefringence of these coffee stain fringes by polarized microscopy and we find the direction of the extraordinary refractive index parallel to the long axis of the nanorods.

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.

[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.

Usefulness of gram staining of blood collected from total parenteral nutrition catheter for rapid diagnosis of catheter-related sepsis.

Science.gov (United States)

Moonens, F; el Alami, S; Van Gossum, A; Struelens, M J; Serruys, E

1994-01-01

The accuracy of Gram staining of blood drawn from catheters used to administer total parenteral nutrition was compared with paired quantitative blood cultures for the diagnosis of catheter-related sepsis. Gram staining was positive in 11 of 18 episodes of catheter-related sepsis documented by quantitative culture (sensitivity, 61%) but in none of the 5 episodes of fever unrelated to catheter infection. Thus, this procedure enabled the rapid presumptive diagnosis and guidance of antimicrobial therapy for total parenteral nutrition catheter sepsis, with a positive predictive value of 100% and a negative predictive value of 42%. PMID:7521359

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 ...

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.

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 ...

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

Science.gov (United States)

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.

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...

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.

Background staining of visualization systems in immunohistochemistry: comparison of the Avidin-Biotin Complex system and the EnVision+ system.

Science.gov (United States)

Vosse, Bettine A H; Seelentag, Walter; Bachmann, Astrid; Bosman, Fred T; Yan, Pu

2007-03-01

The aim of this study was to evaluate specific immunostaining and background staining in formalin-fixed, paraffin-embedded human tissues with the 2 most frequently used immunohistochemical detection systems, Avidin-Biotin-Peroxidase (ABC) and EnVision+. A series of fixed tissues, including breast, colon, kidney, larynx, liver, lung, ovary, pancreas, prostate, stomach, and tonsil, was used in the study. Three monoclonal antibodies, 1 against a nuclear antigen (Ki-67), 1 against a cytoplasmic antigen (cytokeratin), and 1 against a cytoplasmic and membrane-associated antigen and a polyclonal antibody against a nuclear and cytoplasmic antigen (S-100) were selected for these studies. When the ABC system was applied, immunostaining was performed with and without blocking of endogenous avidin-binding activity. The intensity of specific immunostaining and the percentage of stained cells were comparable for the 2 detection systems. The use of ABC caused widespread cytoplasmic and rare nuclear background staining in a variety of normal and tumor cells. A very strong background staining was observed in colon, gastric mucosa, liver, and kidney. Blocking avidin-binding capacity reduced background staining, but complete blocking was difficult to attain. With the EnVision+ system no background staining occurred. Given the efficiency of the detection, equal for both systems or higher with EnVision+, and the significant background problem with ABC, we advocate the routine use of the EnVision+ system.

Design and synthesis of new fluorescent probe for rapid and highly sensitive detection of proteins via electrophoretic gel stain.

Science.gov (United States)

Suzuki, Yoshio; Takagi, Nobuyuki; Chimuro, Tomoyuki; Shinohara, Atsushi; Sakaguchi, Nao; Hiratsuka, Atsunori; Yokoyama, Kenji

2011-06-01

A new fluorescent molecular probe, 2,2'-(1E,1'E)-2,2'-(4-(dicyanomethylene)-4H-pyrane-2,6-diyl)bis(ethene-2,1-diyl)bis(sodium benzenesulfonate) salt (1), possessing the cyanopyranyl moieties and two benzene sulfonic acid groups was designed and synthesized to detect proteins in solution and for high-throughput SDS-PAGE. Compound 1 exhibited no fluorescence in the absence of proteins; however, it exhibited strong fluorescence on the addition of bovine serum albumin as a result of intramolecular charge transfer. Compared with the conventional protocols for in-gel protein staining, such as SYPRO Ruby and silver staining, 1 achieves higher sensitivity, even though it offers a simplified, higher throughput protocol. In fact, the total time required for protein staining was 60-90 min under optimum conditions much shorter than that required by the less-sensitive silver staining or SYPRO Ruby staining protocols. Moreover, 1 was successfully applied to protein identification by mass spectrometry via in-gel tryptic digestion, Western blotting, and native PAGE together with protein staining by 1, which is a modified protocol of blue native PAGE (BN-PAGE). Thus, 1 may facilitate high-sensitivity protein detection, and it may be widely applicable as a convenient tool in various scientific and medical fields. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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....

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...

Cytogenetic data of Partamona peckolti (Hymenoptera, Apidae, Meliponini by C banding and fluorochrome staining with DA/CMA3 and DA/DAPI

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Brito Rute Magalhães

2003-01-01

Full Text Available The stingless bees of the Partamona genus have been studied taxonomically, ecologically and behaviourally, but cytogenetic studies are still rare. The objective of this study was to obtain cytogenetic data to contribute to Partamona peckolti species characterization. Heterochromatin was localized in all chromosome pericentromeric regions but some blocks could be visualized on some large chromosomes arms. A large heterozygous DA-CMA3-positive band was observed on one large chromosome arm, but was completely absent when C banding was applied before fluorochrome staining, with only one small positive band being visualized. Sequential DA-CMA3-NOR staining of interphase nuclei provided coincident positive responses. This suggests that DA-CMA3-positive bands of P. peckolti correspond to nucleolar organizer regions, as previously confirmed for another Partamona species by FISH.

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.

AutoIHC-scoring: a machine learning framework for automated Allred scoring of molecular expression in ER- and PR-stained breast cancer tissue.

Science.gov (United States)

Tewary, S; Arun, I; Ahmed, R; Chatterjee, S; Chakraborty, C

2017-11-01

In prognostic evaluation of breast cancer Immunohistochemical (IHC) markers namely, oestrogen receptor (ER) and progesterone receptor (PR) are widely used. The expert pathologist investigates qualitatively the stained tissue slide under microscope to provide the Allred score; which is clinically used for therapeutic decision making. Such qualitative judgment is time-consuming, tedious and more often suffers from interobserver variability. As a result, it leads to imprecise IHC score for ER and PR. To overcome this, there is an urgent need of developing a reliable and efficient IHC quantifier for high throughput decision making. In view of this, our study aims at developing an automated IHC profiler for quantitative assessment of ER and PR molecular expression from stained tissue images. We propose here to use CMYK colour space for positively and negatively stained cell extraction for proportion score. Also colour features are used for quantitative assessment of intensity scoring among the positively stained cells. Five different machine learning models namely artificial neural network, Naïve Bayes, K-nearest neighbours, decision tree and random forest are considered for learning the colour features using average red, green and blue pixel values of positively stained cell patches. Fifty cases of ER- and PR-stained tissues have been evaluated for validation with the expert pathologist's score. All five models perform adequately where random forest shows the best correlation with the expert's score (Pearson's correlation coefficient = 0.9192). In the proposed approach the average variation of diaminobenzidine (DAB) to nuclear area from the expert's score is found to be 7.58%, as compared to 27.83% for state-of-the-art ImmunoRatio software. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

Comparative study of efficacy, rapidity of detection, and cost-effectiveness of potassium hydroxide, calcofluor white, and Chicago sky blue stains in the diagnosis of dermatophytoses.

Science.gov (United States)

Prakash, R; Prashanth, Hosakere Veerappa; Ragunatha, Shivanna; Kapoor, Meenakshi; Anitha, T K; Krishnamurthy, Veena

2016-04-01

The diagnosis of superficial mycosis such as dermatophytosis is often done clinically. However, in difficult cases, a rapid test with high sensitivity and specificity helps in the immediate confirmation and administration of treatment. The efficacy, rapidity of detection, and cost-effectiveness of KOH preparation, calcofluor white (CW) stain, and Chicago sky blue (CSB) stain in the identification of fungal elements were assessed in patients with dermatophytoses attending the dermatology clinic of a tertiary care hospital. All three tests were performed on each sample collected from 73 patients according to standard procedure. The slides were examined after 5 and 30 minutes in × 10 and × 40 magnifications. The sensitivity and specificity of CW and CSB at 5 and 30 minutes were calculated using KOH preparation as the standard test. CSB stain showed highest positivity (94.5%) within 5 minutes when compared to KOH (75.3%) and CW (83.5%). After 30 minutes, positivity increased in KOH (84.9%) and CW stains (89%), but it remained the same in CSB stain. Both CW and CSB stains when compared to 10% KOH are equally sensitive (100%), but CW was more specific (72.7%), particularly at 30 minutes. When cost of performing tests on 100 specimens is considered, KOH, CW, and CSB stains cost Rs 5, 100, and 15, respectively. CSB stain is a better stain for rapid diagnosis of dermatophytoses because of ease of performance, rapidity of detection, better appreciation of morphology of fungal elements, and cost effectiveness. © 2016 The International Society of Dermatology.

Cytogenetic characterization of Melipona rufiventris Lepeletier 1836 and Melipona mondury Smith 1863 (Hymenoptera, Apidae by C banding and fluorochromes staining

Directory of Open Access Journals (Sweden)

Denilce Meneses Lopes

2008-01-01

Full Text Available The stingless bees Melipona rufiventris and M. mondury were analyzed cytogenetically by conventional staining with Giemsa, C-banding and sequential staining with the fluorochromes CMA3/DA/DAPI. Both species presented 2n = 18 and n = 9, except for one colony of M. rufiventris, in which some individuals had 2n = 19 due to the presence of a B chromosome. After Giemsa staining and C-banding the chromosomes appeared very condensed and presented a high heterochromatic content, making it difficult to localize the centromere and therefore to visualize the chromosomes morphology. The constitutive heterochromatin was located in interstitial chromosome regions covering most of the chromosomes extension and consisted mainly of AT, as shown by DAPI staining. The euchromatin was restricted to the chromosome extremities and was GC-rich, as evidenced by CMA3 staining. The B chromosome was CMA3-negative and DAPI-positive, a heterochromatic constitution similar to that of the A genome chromosomes.

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

Biofilm detection in chronic rhinosinusitis by combined application of hematoxylin-eosin and gram staining.

Science.gov (United States)

Tóth, László; Csomor, Péter; Sziklai, István; Karosi, Tamás

2011-10-01

The pathomechanism of chronic rhinosinusitis with nasal polyposis (CRS/NP) seems to be unclear. Bacterial-, fungal- and combined biofilms might play a potential role in the pathogenesis of various inflammatory diseases and recently in CRS/NP. A prospective, blinded observational study was performed to confirm that the combination of conventional hematoxylin-eosin (HE) and Gram staining protocols could be used to detect bacterial and fungal biofilms in patients with CRS/NP. A total of 50 patients with CRS/NP undergoing endoscopic sinus surgery (ESS) were analyzed. The negative control group consisted of 12 patients undergoing septoplasty for nasal obstruction without CRS/NP. The nasal polyps and inferior turbinate mucosa specimens applied as negative controls were processed to HE and Gram staining. Biofilm was detected in 44 of 50 patients with CRS/NP and in none of 12 negative controls. In our series, HE method showed an obvious correlation with the results of Gram staining and was allocated to be a good predictor of biofilm existence. It was found that the microscopic structure and thickness of biofilms were strongly associated with the integrity of nasal mucosa and with the characteristics of subepithelial cellular infiltration. This study confirmed the presence of bacterial and fungal biofilms on the surface of NPs obtained from patients with CRS. Since biofilms may affect the severity and recurrence rate of CRS treated by ESS they should be detected histologically. In conclusion, HE staining combined with Gram protocol is a robust and reliable method for the detection of bacterial and fungal biofilms in CRS/NP.

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.

Characterization of nonlymphoid cells in rat spleen, with special reference to strongly Ia-positive branched cells in T-cell areas

International Nuclear Information System (INIS)

Dijkstra, C.D.

1982-01-01

By use of a monoclonal antibody against Ia antigen in an immunoperoxidase method, strongly Ia-positive branched cells are found in the T-cell areas of the splenic white pulp of the rat. In order to further characterize these cells, enzyme histochemical characteristics, phagocytic capacity, and irradiation sensitivity have been studied. Evidence is presented that these strongly Ia-positive branched cells represent interdigitating cells. The influence of whole-body irradiation on interdigitating cells is discussed. Comparison with data from the literature on the in vitro dendritic cell isolated from spleen cell suspensions reveals many similarities between the described interdigitating cell in vivo and the dendritic cell in vitro

Gram stain microbiological pattern of upper extremities suppuration at Baptist Medical Centre, Ogbomoso Nigeria: a fifteen month review.

Science.gov (United States)

Oke, A J; Olaolorun, D A; Meier, D E; Tarpley, J L

2011-06-01

Sixty-eight (68) patients with serious upper extremity suppurative infections, presenting within a period of fifteen (15) months, were prospectively studied clinically, Gram stain of aspirates/pus were performed, specimen cultured, planted, and where indicated glucose levels and haemoglobin genotype determined. Half of the patients had hand infections. Staphylococcus aureus was isolated from thirty-nine (39) patients. Gram Negative bacilli, including Salmonella were more isolated from patients with diabetes mellitus or Hgb SS or SC. The Gram stain results correlated with the culture result 90%. When Gram Positive cocci were demonstrated in the primary microscopic examination, cultures were not mandatory. When no organism was demonstrated on primary Gram stain or the patient was diabetic or a sickler, cultures of the specimens were done. The Gram stain, well performed, remains a useful, inexpensive, technologically appropriate laboratory test for abetting decision making in patients with upper extremity suppurative infections. Organisms encountered in this study included: Staphylococcus aureus, Streptococcus pyogenes, Salmonella typhi, Proteus mirabilis, Pseudomonas aeruginosa, and Coliforms.

Mapping stain distribution in pathology slides using whole slide imaging

Directory of Open Access Journals (Sweden)

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.

Tc-99m annexin V imaging and apoptosis staining in rabbit model of osteoarthritis

International Nuclear Information System (INIS)

Kang, Do Young; Jeong, Young Jin; Choi, Sun Mee; Lee, Sung Won; Chung, Won Tae; Yoo, Young Hyun

2004-01-01

Recent studies showed that, in osteoarthritis (OA), articular chondrocytes appeared to be eliminated by apoptosis. Tc-99mm Annexin V has been successfully used for non-invasive gamma imaging of apoptosis in tumor. myocardial infarction and transplantation. We studied Tc-99m Annexin V imaging and apoptosis staining in rabbit model of osteoarthritis. Osteoarthritis was induced in rabbits by intra-articular injection of 1.0 mg collagenase and surgical transection of leg ligament. Animals were dissected at 2, 4, 6 and 8 weeks after the initiation of the injections. For histological observation, the paraffin sections were stained with hematoxylin and eosin. To confirm that osteoarthritis was induced, immunohistochemistry to TRAIL was conducted on the sections and TRAIL positive cells was revealed by DAB. Tc-99m Annexin V was injected 16 ug/1 mCi/kg and regional images were acquired 10 and 60 min postinjection. A few days later Tc-99m MDP imaging was also acquired. Two weeks after injection, the surface layer of cartilage was lost, chondrocytes in the transitional zone have disappeared, and cleft in the transitional zone were shown. Four weeks after injection, moderate cell cloning in transitional and radial zone was appeared. Six weeks after injection, cell cloning was more apparent in the transitional and radial zones. Whereas TRAIL positive cells were not found in the chondrocytes from the control cartilage, most chondrocytes from the collagenase injected cartilage were TRAIL-positive. Tc-99m Annexin V imaging showed increase uptake in knee area of injuried side to normal side. And Tc-99m MDP imaging also had same findings. In rabbit model of osteoarthritis, apoptosis was detected by Tc-99m Annexin V imaging noninvasively and it was correlated by pathologic staining for apoptosis

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.

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.

Hot spot detection for breast cancer in Ki-67 stained slides: image dependent filtering approach

Science.gov (United States)

Niazi, M. Khalid Khan; Downs-Kelly, Erinn; Gurcan, Metin N.

2014-03-01

We present a new method to detect hot spots from breast cancer slides stained for Ki67 expression. It is common practice to use centroid of a nucleus as a surrogate representation of a cell. This often requires the detection of individual nuclei. Once all the nuclei are detected, the hot spots are detected by clustering the centroids. For large size images, nuclei detection is computationally demanding. Instead of detecting the individual nuclei and treating hot spot detection as a clustering problem, we considered hot spot detection as an image filtering problem where positively stained pixels are used to detect hot spots in breast cancer images. The method first segments the Ki-67 positive pixels using the visually meaningful segmentation (VMS) method that we developed earlier. Then, it automatically generates an image dependent filter to generate a density map from the segmented image. The smoothness of the density image simplifies the detection of local maxima. The number of local maxima directly corresponds to the number of hot spots in the breast cancer image. The method was tested on 23 different regions of interest images extracted from 10 different breast cancer slides stained with Ki67. To determine the intra-reader variability, each image was annotated twice for hot spots by a boardcertified pathologist with a two-week interval in between her two readings. A computer-generated hot spot region was considered a true-positive if it agrees with either one of the two annotation sets provided by the pathologist. While the intra-reader variability was 57%, our proposed method can correctly detect hot spots with 81% precision.

Distinction between papillary thyroid hyperplasia and papillary thyroid carcinoma by immunohistochemical staining for cytokeratin 19, galectin-3, and HBME-1.

Science.gov (United States)

Casey, Mary B; Lohse, Christine M; Lloyd, Ricardo V

2003-01-01

The histopathology of papillary thyroid hyperplasia and papillary thyroid carcinoma is similar enough to cause a diagnostic dilemma in a few cases. Both lesions may have papillary fronds with fibrovascular cores, nuclear crowding, and nuclear anisocytosis. Formalin- fixed paraffin-embedded tissues from 30 randomly selected patients with papillary thyroid hyperplasia and an equal number from patients with papillary thyroid carcinoma were analyzed for expression of cytokeratin 19 (CK19), galectin-3, and HBME-1. Cases of papillary thyroid carcinoma had moderate to strong CK19, galectin-3, and HBME-1 reactivity although both CK19 and galectin-3 showed positive staining in a significant number of nonneoplastic thyroid cases. HBME-1 was uncommon in the nonneoplastic cases. These results indicate that HBME-1 may be useful in helping to distinguish papillary thyroid carcinoma from hyperplasia in diagnostically difficult cases.

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...

Comparative Study of Wheatley’s Trichrome Stain and In-vitro Culture against PCR Assay for the Diagnosis of Blastocystis sp. in Stool Samples

Directory of Open Access Journals (Sweden)

Nabilah Amelia MOHAMMAD

2018-03-01

Full Text Available Background: This study evaluated the performance of routine permanent stain and cultivation method in comparison with polymerase chain reaction assay as the reference technique to detect Blastocystis sp.Methods: A cross-sectional study was conducted among aboriginal populations that reside in Pahang, Peninsular Malaysia in Feb to Mar 2015. A total of 359 stool samples were examined using Wheatley’s trichrome stain, in-vitro cultivation in Jones’ medium and PCR assay. Positive amplicons were subjected to sequencing and phylogenetic analysis.Results: Fifty-six (15.6% samples were detected positive with Blastocystis sp. by Wheatley’s trichrome stain and 73 (20.3% by in-vitro culture, while PCR assay detected 71 (19.8% positive samples. Detection rate of Blastocystis sp. was highest in combination of microscopic techniques (27.9%. The sensitivity and specificity of Wheatley’s trichrome staining and in-vitro culture techniques compared to PCR assay were 49.3% (95% CI: 37.2-61.4 and 92.7% (95% CI: 89.1-95.4 and 39.4% (95% CI: 28.0-51.8 and 84.4% (95% CI: 79.7-88.4, respectively. However, the sensitivity [60.6% (95% CI: 48.3-71.9] of the method increased when both microscopic techniques were performed together. False negative results produced by microscopic techniques were associated with subtype 3. The agreement between Wheatley’s trichrome stain, in-vitro culture and combination of microscopic techniques with PCR assay were statistically significant by Kappa statistics (Wheatley’s trichrome stain: K = 0.456, P<0.001; in-vitro culture: K = 0.236, P<0.001 and combination techniques: K = 0.353, P<0.001.Conclusion: The combination of microscopic technique is highly recommended to be used as a screening method for the diagnosis of Blastocystis infection either for clinical or epidemiological study to ensure better and accurate diagnosis.

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.

Nucleic acid stains as indicators of Giardia muris viability following cyst inactivation.

Science.gov (United States)

Taghi-Kilani, R; Gyürék, L L; Millard, P J; Finch, G R; Belosevic, M

1996-06-01

A reliable viability assay for Giardia is required for the development of disinfection process design criteria and pathogen monitoring by water treatment utilities. Surveys of single-staining nucleic acid dyes (stain dead parasites only), and double-staining vital dye kits from Molecular Probes (stain live and dead parasites) were conducted to assess the viability of untreated, heat-killed, and chemically inactivated Giardia muris cysts. Nucleic acid staining results were compared to those of in vitro excystation and animal infectivity. Nucleic acid stain, designated as SYTO-9, was considered the best among the single-staining dyes for its ability to stain dead cysts brightly and its relatively slow decay rate of visible light emission following DNA binding. SYTO-9 staining was correlated to animal infectivity. A Live/Dead BacLight was found to be the better of 2 double-staining viability kits tested. Logarithmic survival ratios based on SYTO-9 and Live/Dead BacLight were compared to excystation and infectivity results for G. muris cysts exposed to ozone or free chlorine. The results indicate that SYTO-9 and Live/Dead BacLight staining is stable following treatment of cysts with chemical disinfectants.

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.

Start Position Strongly Influences Fixation Patterns during Face Processing: Difficulties with Eye Movements as a Measure of Information Use

Science.gov (United States)

Arizpe, Joseph; Kravitz, Dwight J.; Yovel, Galit; Baker, Chris I.

2012-01-01

Fixation patterns are thought to reflect cognitive processing and, thus, index the most informative stimulus features for task performance. During face recognition, initial fixations to the center of the nose have been taken to indicate this location is optimal for information extraction. However, the use of fixations as a marker for information use rests on the assumption that fixation patterns are predominantly determined by stimulus and task, despite the fact that fixations are also influenced by visuo-motor factors. Here, we tested the effect of starting position on fixation patterns during a face recognition task with upright and inverted faces. While we observed differences in fixations between upright and inverted faces, likely reflecting differences in cognitive processing, there was also a strong effect of start position. Over the first five saccades, fixation patterns across start positions were only coarsely similar, with most fixations around the eyes. Importantly, however, the precise fixation pattern was highly dependent on start position with a strong tendency toward facial features furthest from the start position. For example, the often-reported tendency toward the left over right eye was reversed for the left starting position. Further, delayed initial saccades for central versus peripheral start positions suggest greater information processing prior to the initial saccade, highlighting the experimental bias introduced by the commonly used center start position. Finally, the precise effect of face inversion on fixation patterns was also dependent on start position. These results demonstrate the importance of a non-stimulus, non-task factor in determining fixation patterns. The patterns observed likely reflect a complex combination of visuo-motor effects and simple sampling strategies as well as cognitive factors. These different factors are very difficult to tease apart and therefore great caution must be applied when interpreting absolute

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.

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.

Chromosome-specific staining to detect genetic rearrangements

Energy Technology Data Exchange (ETDEWEB)

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

2013-04-09

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

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...

Cancer-associated fibroblasts are positively correlated with metastatic potential of human gastric cancers

Directory of Open Access Journals (Sweden)

Zhang Hao

2010-06-01

Full Text Available Abstract Background The prognosis of gastric cancer patients is difficult to predict because of defects in establishing the surgical-pathological features. Cancer-associated fibroblasts (CAFs have been found to play prominent role in promoting tumor growth, invasion and metastasis. Thus raises the hypothesis that the extent of CAFs prevalence may help to establish the prognosis of gastric cancer patients. Methods Immunochemistry and realtime-PCR experiments were carried out to compare the expression of proteins which are specific markers of CAFs or secreted by CAFs in the tumor and normal tissue specimens. The extent of CAFs' prevalence was graded according to immunochemical staining, and correlation was further analyzed between CAFs' prevalence and other tumor characteristics which may influence the prognosis of gastric cancer patients. Results Nearly 80 percent of normal gastric tissues were negative or weak positive for CAFs staining, while more than 60 percent of gastric cancer tissues were moderate or strong positive for CAFs staining. Realtime-PCR results also showed significant elevated expression of FAP, SDF-1 and TGF-β1 in gastric cancer tissues compared to normal gastric tissues. Further analysis showed that CAFs' prevalence was correlated with tumor size, depth of the tumor, lymph node metastasis, liver metastasis or peritoneum metastasis. Conclusions Reactive cancer associated fibroblasts (CAFs were frequently accumulated in gastric cancer tissues, and the prevalence of CAFs was correlated with tumor size, depth of the tumor and tumor metastasis, thus give some supports for establishing the prognosis of the gastric cancer patients.

A molecular gram stain using broad range PCR and pyrosequencing technology: a potentially useful tool for diagnosing orthopaedic infections.

Science.gov (United States)

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

2005-06-01

The bacteria associated with orthopaedic infections are usually common gram-positive and gram-negative bacteria. This fundamental grouping of bacteria is a necessary first step in the selection of appropriate antibiotics. Since polymerase chain reaction (PCR) is more rapid and may be more sensitive than culture, we developed a postamplification pyrosequencing method to subcategorize bacteria based on a few nucleotide polymorphisms in the 16S rRNA gene. We validated this method using well-characterized strains of bacteria and applied it to specimens from spinal surgery cases with suspected infections. Lysates of 114 bacteria including 75 species were created following standard cultivation to obtain DNA. The DNA was amplified by a broad-range real-time PCR. The amplicons were evaluated by pyrosequencing and were classified as gram-positive, gram-negative, or acid-fast bacilli based on the first three to five nucleotides sequenced. In addition, clinical cases of suspected infection were obtained from spinal surgery. The results of the "molecular Gram stain" were compared with the results of traditional Gram stain and culture. The lysates of 107 (93.9%) of the bacteria extracts tested were appropriately categorized as gram-positive and gram-negative or as acid-fast bacilli on the basis of this assay. The sensitivity and specificity of this assay were 100% and 97.4% for gram-positive and 88.3% and 100% for gram-negative isolates. All of the five clinical samples were appropriately categorized as containing gram-positive or gram-negative bacteria with this assay. This study demonstrates that high sensitivity and specificity of a molecular gram stain may be achieved using broad-range real-time PCR and pyrosequencing.

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.

Performance of Gram staining on blood cultures flagged negative by an automated blood culture system.

Science.gov (United States)

Peretz, A; Isakovich, N; Pastukh, N; Koifman, A; Glyatman, T; Brodsky, D

2015-08-01

Blood is one of the most important specimens sent to a microbiology laboratory for culture. Most blood cultures are incubated for 5-7 days, except in cases where there is a suspicion of infection caused by microorganisms that proliferate slowly, or infections expressed by a small number of bacteria in the bloodstream. Therefore, at the end of incubation, misidentification of positive cultures and false-negative results are a real possibility. The aim of this work was to perform a confirmation by Gram staining of the lack of any microorganisms in blood cultures that were identified as negative by the BACTEC™ FX system at the end of incubation. All bottles defined as negative by the BACTEC FX system were Gram-stained using an automatic device and inoculated on solid growth media. In our work, 15 cultures that were defined as negative by the BACTEC FX system at the end of the incubation were found to contain microorganisms when Gram-stained. The main characteristic of most bacteria and fungi growing in the culture bottles that were defined as negative was slow growth. This finding raises a problematic issue concerning the need to perform Gram staining of all blood cultures, which could overload the routine laboratory work, especially laboratories serving large medical centers and receiving a large number of blood cultures.

Colorimetric sensor for triphosphates and their application as a viable staining agent for prokaryotes and eukaryotes.

Science.gov (United States)

Ghosh, Amrita; Shrivastav, Anupama; Jose, D Amilan; Mishra, Sanjiv K; Chandrakanth, C K; Mishra, Sandhya; Das, Amitava

2008-07-15

The chromogenic complex 1 x Zn (where 1 is (E)-4-(4-dimethylamino-phenylazo)-N,N-bispyridin-2-ylmethyl-benzenesulfonamide) showed high affinity toward the phosphate ion in tetrabutylammonium phosphate in acetonitrile solution and could preferentially bind to adenosine triphosphate (ATP) in aqueous solution at physiological pH. This binding caused a visual change in color, whereas no such change was noticed with other related anions (adenosine monophosphate, adenosine diphosphate, pyrophosphate, and phosphate) of biological significance. Thus, 1 x Zn could be used as a staining agent for different biological cells through binding to the ATP, generated in situ by the mitochondria (in eukaryotes). For prokaryotes (bacteria) the cell membrane takes care of the cells' energy conversion, since they lack mitochondria. ATP is produced in their unique cell structure on the cell membrane, which is not found in any eukaryotes. These stained cells could be viewed with normal light microscopy. This reagent could even be used for distinguishing the gram-positive and the gram-negative bacteria (prokaryotes). This dye was found to be nonlipophilic in nature and nontoxic to living microbes (eukaryotes and prokaryotes). Further, stained cells were found to grow in their respective media, and this confirmed the maintenance of viability of the microbes even after staining, unlike with many other dyes available commercially.

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...

Determining the Effect of Calculus, Hypocalcification, and Stain on Using Optical Coherence Tomography and Polarized Raman Spectroscopy for Detecting White Spot Lesions

Directory of Open Access Journals (Sweden)

Amanda Huminicki

2010-01-01

Full Text Available Optical coherence tomography (OCT and polarized Raman spectroscopy (PRS have been shown as useful methods for distinguishing sound enamel from carious lesions ex vivo. However, factors in the oral environment such as calculus, hypocalcification, and stain could lead to false-positive results. OCT and PRS were used to investigate extracted human teeth clinically examined for sound enamel, white spot lesion (WSL, calculus, hypocalcification, and stain to determine whether these factors would confound WSL detection with these optical methods. Results indicate that OCT allowed differentiating caries from sound enamel, hypocalcification, and stain, with calculus deposits recognizable on OCT images. ANOVA and post-hoc unequal N HSD analyses to compare the mean Raman depolarization ratios from the various groups showed that the mean values were statistically significant at P<.05, except for several comparison pairs. With the current PRS analysis method, the mean depolarization ratios of stained enamel and caries are not significantly different due to the sloping background in the stained enamel spectra. Overall, calculus and hypocalcification are not confounding factors affecting WSL detection using OCT and PRS. Stain does not influence WSL detection with OCT. Improved PRS analysis methods are needed to differentiate carious from stained enamel.

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.

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

Utility of Fite-Faraco stain for both mast cell count and bacillary index in skin biopsies of leprosy patients.

Science.gov (United States)

Chatura, K R; Sangeetha, S

2012-01-01

To assess the utility of a single stain for both mast cell count and bacillary index (BI), 50 skin-biopsie patients were stained with Fite-Faraco (FF) stain, viewed under oil immersion and BI calculated using the Ridley's logarithmic scale, and mast cells counted as the number of cells per mm2. Mean mast cell count per mm2 at the tuberculoid pole was lowest in TT 7.9 and highest in BT 14.23. At the lepromatous end, it was highest in BL 9.21, while in LL it was 8.23. Highest counts were seen in the borderline types overall. The correlation coefficient between histopathological diagnosis and BI is 0.822 which is a positive correlation to a significant degree. The correlation coefficient between histopathological diagnosis and mast cell count was found to be -0.17, which is a negative correlation but not to a significant degree. FF stain was utilised to visualise both bacilli for estimation of BI and mast cells for mast cell count, a seldom attempted feature in literature.

Contribution of immunocytochemical stainings for galectin-3, CD44, and HBME1 to fine-needle aspiration cytology diagnosis of papillary thyroid carcinoma.

Science.gov (United States)

Das, Dilip K; Al-Waheeb, Salah K M; George, Sara S; Haji, Bahiyah I; Mallik, Mrinmay K

2014-06-01

In cytology practice some papillary thyroid carcinoma (PTC) cases have indeterminate diagnoses and overlapping cytological features with benign lesions. This study was undertaken to find out if immunocytochemistry using Galectin-3, CD-44 and HBME-1 could be of help in such situations. Forty-six cases consisting of 22 malignancy (PTC) cases, 7 suspicious of (S/O) PTC, 1 follicular neoplasm, 5 follicular lesion of undetermined significance (FLUS), and 11 benign (colloid goiter) cases diagnosed by FNA were included in this study. Staining reactions were graded in a sliding scale of -, 1+, 2+, 3+, and 4+. In an assessment of 100 cells, each cell with weak, and moderate to strong positive reaction were assigned a score of 1 and 4, respectively. Staining reaction of ≥+2 and scores >100 were considered positive. Frequency of cases with ≥+2 reaction, and scores >100 for each of Galectin-3, CD-44, and HBME-1 were significantly higher in PTC or combined PTC and S/O PTC cases as compared with FLUS and benign cases taken together (P = 0.01744 to 0.00000). When the cases were compared according to histological malignant and benign diagnoses, the difference was also significant in respect of ≥+2 reaction, and scores >100 for Galectin-3 and CD44 (P = 0.04923 to 0.00947); however, there was no significant difference, when these parameters for HBME1 were compared. Galectin 3, CD 44, and to some extent HBME 1 are useful immunocytochemical parameters with potential to support FNAC diagnosis of PTC, especially in situations with difficult differential diagnoses. Copyright © 2013 Wiley Periodicals, Inc.

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

Características morfo-tintoriales en el ciclo celular de Acinetobacter baumannii por los métodos de Gram y 4’, 6-diamidino-2’-fenilindol, dihidrocloruro | Morphological and staining characteristics in cell cycle of Acinetobacter baumannii by the methods of Gram and 4´, 6-diamidino-2´-phenylindole, dihydrochloride

Directory of Open Access Journals (Sweden)

Evelin Flores Hernández

2017-11-01

Full Text Available Several studies have reported variations in bacterial staining by Gram, which are associated with the content of nucleic acids in the bacterial cell cycle. In order to give continuity to the microscopical studies pertaining to the bacterial cell cycle, the morphological and staining characteristics were evaluated by Gram and 4', 6-diamidino-2'-phenylindole, dihydrochloride (DAPI in the cell cycle of Acinetobacter baumannii. From the synchronic cultures of 4 strains of A. baumannii, the cell cycle was studied incubating them in aerobiosis at 30ºC, taking aliquots every 5 min for 2 h. In turn, the respective extensions were made and stained with Gram and DAPI for their posterior microscopical study. In the statistical analysis Fischer's test was used to associate morphological and the staining characteristics by Gram and DAPI. In 74% of cycle times, the cocoid form predominated and 67% in the negative Gram staining. When determining the mean and standard deviation of bacterial morphological values, it was observed that between 15 and 55 min the bacterium initiates a new cell cycle, decreasing its diameter and length. There was a statistically significant association (p < 0.05 between the lax and compact appearance of the material with the light and intense fluorescent, respectively; likewise, between the strong Gram positive character and the intense fluorescence. It is concluded that in the Gram positive reaction, the DNA is possibly involved and that the purple material of compact and lax aspect observed by Gram staining in the bacterial cytoplasm correspond to the nucleoid and the morphological changes obey to the cellular growth process. These findings contribute to explain the mechanism of Gram staining, as well as its variability and the pleomorphism observed in this bacterial genus.

Newly formed skeletal muscle fibers are prone to false positive immunostaining by rabbit antibodies

DEFF Research Database (Denmark)

Andersen, Ditte C; Kliem, Anette; Schrøder, Henrik Daa

2011-01-01

rely on controls that reveal non-specific binding by the secondary antibody and neglect that the primary rabbit antibody itself may cause false positive staining of the muscle. We suggest that reliable immuno-based protein detection in newly formed muscle fibers at least requires a nonsense rabbit......Reports on muscle biology and regeneration often implicate immuno(cyto/histo)chemical protein characterization using rabbit polyclonal antibodies. In this study we demonstrate that newly formed myofibers are especially prone to false positive staining by rabbit antibodies and this unwanted staining...

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...

Application of sol-gel based sensors to environmental monitoring of Maumejean stained glass windows housed in two different buildings at downtown Madrid

Energy Technology Data Exchange (ETDEWEB)

Pena-Poza, J.; Conde, J. F.; Agua, F.; Garcia-Heras, M.; Villegas, M. A.

2013-07-01

Degradation of historical stained glass windows is mainly caused by acid attack enhanced by humidity and pollutants. Accordingly, their preventive conservation should include environmental evaluation. Maumejean's stained glass windows (c 1940) of two buildings located at downtown Madrid have been monitored by sol-gel sensors of acidity and temperature. The philosophy was the application of innovative glassy sol-gel sensors to assess the conservation conditions of stained glass windows, i.e. modern materials for preservation of historical materials. Conservation conditions (environmental acidity and temperature) of restored and non-restored stained glass windows have been recorded throughout 13 months. The main contributing parameter to outdoor acidity is proximity to road traffic, which produces acid species able to diminish two units of pH with respect to neutral conditions. This acid environment affects both sides of stained glass windows, even in those protected with a glazing system, which allows natural ventilation. Other contributing parameters to increase the air acidity were facade orientation, sensor position, distance to pollutants sources, human interaction and uncontrolled ventilation. (Author)

Comparative analysis of Gram's stain, PNA-FISH and Sepsityper with MALDI-TOF MS for the identification of yeast direct from positive blood cultures.

Science.gov (United States)

Gorton, Rebecca L; Ramnarain, P; Barker, K; Stone, N; Rattenbury, S; McHugh, T D; Kibbler, C C

2014-10-01

Fungaemia diagnosis could be improved by reducing the time to identification of yeast from blood cultures. This study aimed to evaluate three rapid methods for the identification of yeast direct from blood cultures; Gram's stain analysis, the AdvanDX Peptide Nucleic Acid in Situ Hybridisation Yeast Traffic Light system (PNA-FISH YTL) and Bruker Sepsityper alongside matrix-assisted laser desorption ionisation time of flight mass spectrometry (MALDI-TOF MS). Fifty blood cultures spiked with a known single yeast strain were analysed by blinded operators experienced in each method. Identifications were compared with MALDI-TOF MS CHROMagar Candida culture and ITS rRNA sequence-based identifications. On first attempt, success rates of 96% (48/50) and 76% (36/50) were achieved using PNA-FISH YTL and Gram's stain respectively. MALDI-TOF MS demonstrated a success rate of 56% (28/50) when applying manufacturer's species log score thresholds and 76% (38/50) using in-house parameters, including lowering the species log score threshold to >1.5. In conclusion, PNA-FISH YTL demonstrated a high success rate successfully identifying yeast commonly encountered in fungaemia. Sepsityper(™) with MALDI-TOF MS was accurate but increased sensitivity is required. Due to the misidentification of commonly encountered yeast Gram's stain analysis demonstrated limited utility in this setting. © 2014 Blackwell Verlag GmbH.

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.

Differentiation of Body Fluid Stains on Fabrics Using External Reflection Fourier Transform Infrared Spectroscopy (FT-IR) and Chemometrics.

Science.gov (United States)

Zapata, Félix; de la Ossa, Ma Ángeles Fernández; García-Ruiz, Carmen

2016-04-01

Body fluids are evidence of great forensic interest due to the DNA extracted from them, which allows genetic identification of people. This study focuses on the discrimination among semen, vaginal fluid, and urine stains (main fluids in sexual crimes) placed on different colored cotton fabrics by external reflection Fourier transform infrared spectroscopy (FT-IR) combined with chemometrics. Semen-vaginal fluid mixtures and potential false positive substances commonly found in daily life such as soaps, milk, juices, and lotions were also studied. Results demonstrated that the IR spectral signature obtained for each body fluid allowed its identification and the correct classification of unknown stains by means of principal component analysis (PCA) and soft independent modeling of class analogy (SIMCA). Interestingly, results proved that these IR spectra did not show any bands due to the color of the fabric and no substance of those present in daily life which were analyzed, provided a false positive. © The Author(s) 2016.

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.

Classification of phytoplankton cells as live or dead using the vital stains fluorescein diacetate and 5-chloromethylfluorescein diacetate.

Science.gov (United States)

MacIntyre, Hugh L; Cullen, John J

2016-08-01

Regulations for ballast water treatment specify limits on the concentrations of living cells in discharge water. The vital stains fluorescein diacetate (FDA) and 5-chloromethylfluorescein diacetate (CMFDA) in combination have been recommended for use in verification of ballast water treatment technology. We tested the effectiveness of FDA and CMFDA, singly and in combination, in discriminating between living and heat-killed populations of 24 species of phytoplankton from seven divisions, verifying with quantitative growth assays that uniformly live and dead populations were compared. The diagnostic signal, per-cell fluorescence intensity, was measured by flow cytometry and alternate discriminatory thresholds were defined statistically from the frequency distributions of the dead or living cells. Species were clustered by staining patterns: for four species, the staining of live versus dead cells was distinct, and live-dead classification was essentially error free. But overlap between the frequency distributions of living and heat-killed cells in the other taxa led to unavoidable errors, well in excess of 20% in many. In 4 very weakly staining taxa, the mean fluorescence intensity in the heat-killed cells was higher than that of the living cells, which is inconsistent with the assumptions of the method. Applying the criteria of ≤5% false negative plus ≤5% false positive errors, and no significant loss of cells due to staining, FDA and FDA+CMFDA gave acceptably accurate results for only 8-10 of 24 species (i.e., 33%-42%). CMFDA was the least effective stain and its addition to FDA did not improve the performance of FDA alone. © 2016 The Authors. Journal of Phycology published by Wiley Periodicals, Inc. on behalf of Phycological Society of America.

Damping at positive frequencies in the limit J⊥-->0 in the strongly correlated Hubbard model

Science.gov (United States)

Mohan, Minette M.

1992-08-01

I show damping in the two-dimensional strongly correlated Hubbard model within the retraceable-path approximation, using an expansion around dominant poles for the self-energy. The damping half-width ~J2/3z occurs only at positive frequencies ω>5/2Jz, the excitation energy of a pure ``string'' state of length one, where Jz is the Ising part of the superexchange interaction, and occurs even in the absence of spin-flip terms ~J⊥ in contrast to other theoretical treatments. The dispersion relation for both damped and undamped peaks near the upper band edge is found and is shown to have lost the simple J2/3z dependence characteristic of the peaks near the lower band edge. The position of the first three peaks near the upper band edge agrees well with numerical simulations on the t-J model. The weight of the undamped peaks near the upper band edge is ~J4/3z, contrasting with Jz for the weight near the lower band edge.

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.

The utility of periodic acid schiff with diastase and alcian blue stains on fine needle aspirates of breast and salivary gland neoplasms

Directory of Open Access Journals (Sweden)

N K Panicker

2012-01-01

Conclusion : Intracytoplasmic PAS-D-positive globules may be useful in differentiating benign and malignant lesions of breast. The presence of PAS-D positive granules are useful in differentiating various lesions of salivary glands. AB staining of stromal fragments in pleomorphic adenoma is useful in differentiating it from basal cell adenoma.

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.

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

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

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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.

Colposcopic characteristics and Lugol׳s staining differentiate anal high-grade and low-grade squamous intraepithelial lesions during high resolution anoscopy

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Naomi Jay

2015-12-01

Full Text Available Background: Anal squamous intraepithelial lesions (SIL and cancers are increased in immunocompromised populations. Based upon anatomic and histologic similarities, the cervix is used as the model for anal screening. During cervical colposcopy, acetic acid (AA and Lugol׳s staining (LS result in characteristic changes that help distinguish low-grade (LSIL from high-grade (HSIL. Lesion characteristics were evaluated for their ability to distinguish anal (aLSIL from anal (aHSIL during high-resolution anoscopy after application of AA and LS. Methods: AA-stained lesions were described using standard cervical colposcopic criteria. LS was then applied and lesions were characterized as Lugol׳s-negative (L−, Lugol׳s-partial (L+/−, or Lugol׳s positive (L+ and then biopsied. Biopsies were characterized as benign, squamous atypia, LSIL or HSIL. Results: 835 anal lesions were analyzed. Sensitivity and positive predictive value (PPV for aHSIL were highest for characteristics associated with cervical (cHSIL. L− was independently associated with aHSIL (OR=4.7, 95% CI=3.4–6.7. In multiple logistic regression analysis, significant predictors of aHSIL were flat contour (OR=2.24, 95% CI=1.3–3.8, mosaic pattern (OR=2.0, 95% CI=1.4–2.9, vascular punctation (OR=1.5, 95% CI=1.1–2.1 and L− (OR=2.3, 95% CI=1.5–3.4. L− staining improved the PPV of aHSIL almost twofold in lesions that otherwise had a colposcopic impression of LSIL. Conclusions: Evaluating acetowhite lesions for contour, surface, vascularity, and LS may maximize the likelihood of identifying aHSIL. Keywords: Anal, HSIL, High resolution anoscopy, Lugol׳s staining

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

Detection of Cryptosporidium sp infection by PCR and modified acid fast staining from potassium dichromate preserved stool

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Agnes Kurniawan

2009-09-01

Full Text Available Aim To identify the frequency of Cryptosporidium infection in children below 3 years old by examining concentrated long term preserved stool using PCR detection of 18S rRNA gene and compared with modified acid fast staining technique.Methods Hundred eighty eight stools from children ≤ 3 years old were stored for 13 months in 2.5% K2Cr2O7 solution at 40C. Cryptosporidium oocysts were isolated by water-ether concentration technique. The concentrates were smeared onto object glass and stained with modified acid fast staining, and the rest of the concentrates were DNA extracted by freezing and thawing cycles and proteinase K digestion, then direct PCR was done to detect 18S rRNA gene.Result The proportion of positive stools for Cryptosporidium sp by acid fast staining from concentrated stools and 18S rRNA PCR were 4.8% and 34.6% respectively, which showed statistically significant difference.Conclusion The frequency of Cryptosporidium infection among children ≤ 3 years old was very high and stool storage in K2Cr2O7 for 13 months did not affect the PCR result. High prevalence of Cryptosporidium infection indicated high transmission in that area and the potential to be transmitted to other individuals such as the immunocompromised. (Med J Indones 2009;18:147-52Key words: 18S rRNA, cryptosporidiosis

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 ...

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

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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

Performing Gram stain directly on catheter tips: assessment of the quality of the observation process.

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Guembe, M; Pérez-Granda, M J; Rivera, M L; Martín-Rabadán, P; Bouza, E

2015-06-01

A previous study performed in our institution showed that catheter tip (CT) staining by combining acridine orange and Gram stain (GS) before culture anticipated catheter colonization with exhaustive and careful observation by a highly trained technician. Our objective was to assess the validity values of GS without acridine orange on an external smear of CT for predicting catheter colonization and catheter-related bloodstream infection (C-RBSI). We compared different periods of observation and the results of two technicians with different levels of professional experience. Over a 5-month period, the roll-plate technique was preceded by direct GS of all CTs sent to the microbiology laboratory. The reading was taken at ×100 by two observers with different skill levels. Each observer performed a routine examination (3 min along three longitudinal lines) and an exhaustive examination (5 min along five longitudinal lines). The presence of at least one cell was considered positive. All slides were read before culture results were known. We included a total of 271 CTs from 209 patients. The prevalence of catheter colonization and C-RBSI was 16.2 % and 5.1 %, respectively. Routine and exhaustive examinations revealed only 29.5 % and 40.9 % of colonized catheters, respectively (p staining is performed exhaustively. However, the decision to implement this approach in daily routine will depend on the prevalence rate of catheter colonization at each institution.

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.

Epicardial adipose tissue volume and adipocytokine imbalance are strongly linked to human coronary atherosclerosis.

Science.gov (United States)

Shimabukuro, Michio; Hirata, Yoichiro; Tabata, Minoru; Dagvasumberel, Munkhbaatar; Sato, Hiromi; Kurobe, Hirotsugu; Fukuda, Daiju; Soeki, Takeshi; Kitagawa, Tetsuya; Takanashi, Shuichiro; Sata, Masataka

2013-05-01

The impact of epicardial adipose tissue (EAT) over abdominal or overall adiposity on coronary artery disease (CAD) is currently unknown. We compared the association among EAT volume (EATV), cytokine/adipocytokine profiles in EAT and subcutaneous fat, and atherogenic CAD. Paired samples were obtained from EAT and subcutaneous adipose tissue during elective cardiac surgery for CAD (n=50) or non-CAD (n=50). EATV was the sum of cross-sectional EAT areas, and visceral and subcutaneous fat areas were determined at the umbilicus level on computed tomography scans. CD68(+), CD11c(+), and CD206(+) cells were counted using immunohistochemical staining. Cytokine/adipocytokine expression was evaluated using quantitative real-time polymerase chain reaction. Multivariate analysis indicated that male sex, age, diabetes mellitus, high triglycerides, and low high-density lipoprotein cholesterol, and EATV index (EATV/body surface area, cm(3)/m(2)) were significant CAD predictors (corrected R(2)=0.401; PEATV index positively correlated with the CD68(+) and CD11c(+) cell numbers and nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3), interleukin-1β, and interleukin-1R expression; and negatively correlated with adiponectin expression in EAT. A multivariate analysis model, including CD68(+) cells and interleukin-1β, and adiponectin expression in EAT strongly predicted CAD (corrected R(2)=0.756; PEATV and macrophage and cytokine/adipocytokine signals in EAT strongly correlated with CAD. Our findings suggest that EATV and adipocytokine imbalance are strongly linked to human coronary atherosclerosis.

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

Studies of the endothelial origin of cells in systemic angioendotheliomatosis and other vascular lesions of the brain and meninges using ulex europaeus lectin stains.

Science.gov (United States)

Schelper, R L; Olson, S P; Carroll, T J; Hart, M N; Witters, E

1986-01-01

Ulex europaeus agglutinin I (UEA-I) is a plant lectin which binds specifically to alpha-L-fucose moieties on the surface glycoproteins of human endothelial cells. The binding is completely inhibited by preincubation of the lectin with fucose. UEA-I can be conjugated directly to fluorescein or peroxidase and can be used to stain endothelium of paraffin embedded tissues. UEA-I staining was evaluated on normal and infarcted brain, systemic angioendotheliomatosis, metastatic epidural angiosarcoma, hemangioendothelioma, hemangioblastoma, angioblastic meningioma of both the hemangioblastic and hemangiopericytic types, and vascular meningioma. The endothelium, but not neuropil of normal and infarcted brain was positive for UEA-I. The tumor cells of hemangioendothelioma and angiosarcoma also stained. However, no staining was seen in malignant intravascular cells of angioendotheliomatosis, the stromal cells of hemangioblastoma, or pericytes of angioblastic meningioma. It is concluded that the malignant cells in angioendotheliomatosis, the stromal cells of hemangioblastoma and the pericytes of angioblastic meningioma do not produce surface glycoproteins characteristic of endothelial cells.

Small Winter Thunderstorm with Sprites and Strong Positive Discharge

Science.gov (United States)

Suzuki, Tomoyuki; Hayakawa, Masashi; Michimoto, Koichiro

A sprite campaign was conducted in the Hokuriku area of Japan during a winter of 2004/2005. On the basis of a combined analysis of the data from various instruments (CCD cameras, radar, VHF/LF∼MF lightning mapping system, field mill network, and ELF detector), we studied meteorological and electrical structures for winter thunderstorms and sprite-producing positive discharge. Typical winter sprite parent thunderstorms had a meso-scale cloud area with embedded small convective cells. Some small winter thunderstorms accompanied by the most frequent sprite events were found to cause 2∼3 sprite events during a short interval of about 3∼5 min. When the sprites were observed, the extent of the convective cells at 20 dBZ counter was atmost ∼20 × 20 km. The VHF sources associated with sprites were located near south of the convective cell and were mapped within very small areas of at most ∼10 × 10 km. This fact shows that some small winter thunderstorms can generate large positive charge associated with sprites. We will present the analysis of such a small thunderstorms with sprites and positive lightning discharges.

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

Science.gov (United States)

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

Neuronal density, size and shape in the human anterior cingulate cortex: a comparison of Nissl and NeuN staining.

Science.gov (United States)

Gittins, Rebecca; Harrison, Paul J

2004-03-15

There are an increasing number of quantitative morphometric studies of the human cerebral cortex, especially as part of comparative investigations of major psychiatric disorders. In this context, the present study had two aims. First, to provide quantitative data regarding key neuronal morphometric parameters in the anterior cingulate cortex. Second, to compare the results of conventional Nissl staining with those observed after immunostaining with NeuN, an antibody becoming widely used as a selective neuronal marker. We stained adjacent sections of area 24b from 16 adult brains with cresyl violet or NeuN. We measured the density of pyramidal and non-pyramidal neurons, and the size and shape of pyramidal neurons, in laminae II, III, Va, Vb and VI, using two-dimensional counting methods. Strong correlations between the two modes of staining were seen for all variables. However, NeuN gave slightly higher estimates of neuronal density and size, and a more circular perikaryal shape. Brain pH was correlated with neuronal size, measured with both methods, and with neuronal shape. Age and post-mortem interval showed no correlations with any parameter. These data confirm the value of NeuN as a tool for quantitative neuronal morphometric studies in routinely processed human brain tissue. Absolute values are highly correlated between NeuN and cresyl violet stains, but cannot be interchanged. NeuN may be particularly useful when it is important to distinguish small neurons from glia, such as in cytoarchitectural studies of the cerebral cortex in depression and schizophrenia.

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

Science.gov (United States)

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.

Histological identification of H. pylori stained by hematoxylin-eosin and Giemsa: review for quality control

Directory of Open Access Journals (Sweden)

Marcela S. Boldt

2015-04-01

Full Text Available Introduction: Several special staining methods are available for H. pylori (Hp identification in histological sections of chronic gastritis (CG, including the routine hematoxylin-eosin (HE method. Some reports suggest that ancillary stains are not always needed to establish the diagnosis of Hp infection. In addition, the benefit of using them, when biopsies show minimal inflammation, is not clear. Objective: We performed a retrospective study to compare the usefulness of HE with Giemsa method for the histopathological diagnosis of Hp in tissue sections. Methods: Histological sections from 390 consecutive patients were reviewed. The patients were registered in the histopathology laboratory of Instituto Alfa de Gastroenterologia, Hospital das Clínicas da Universidade Federal de Minas Gerais (UFMG, Belo Horizonte, Brasil. They were divided in 4 groups according to the gastric inflammatory changes as follows: Group I, gastric mucosa with normal morphology or minimal inflammatory changes (n = 146; Group II, chronic gastritis (CG with mild inflammatory activity (n = 101; Group III, CG with patent inflammatory activity (n = 123; Group IV, patients with atrophic body gastritis (n = 20. All histological sections were carefully evaluated by 2 examiners at the oil immersion objective (1000×. Results: The identification of Hp was positive by Giemsa and HE, respectively at: Group III, 111 (90.2% and 93 (75.6% patients (p < 0.01; Group II, 43 (42.6% and 29 (28.7% patients (p < 0.05. Hp was negative in Groups I and IV. Conclusion: The results show that Giemsa stain is superior to HE for histological identification of Hp in CG. Although Hp could be identified by HE stain in the majority of CG cases, a significant number of infected patients may be neglected, regardless the intensity of the inflammatory response.

Identification of immune cell infiltration in hematoxylin-eosin stained breast cancer samples: texture-based classification of tissue morphologies

Science.gov (United States)

Turkki, Riku; Linder, Nina; Kovanen, Panu E.; Pellinen, Teijo; Lundin, Johan

2016-03-01

The characteristics of immune cells in the tumor microenvironment of breast cancer capture clinically important information. Despite the heterogeneity of tumor-infiltrating immune cells, it has been shown that the degree of infiltration assessed by visual evaluation of hematoxylin-eosin (H and E) stained samples has prognostic and possibly predictive value. However, quantification of the infiltration in H and E-stained tissue samples is currently dependent on visual scoring by an expert. Computer vision enables automated characterization of the components of the tumor microenvironment, and texture-based methods have successfully been used to discriminate between different tissue morphologies and cell phenotypes. In this study, we evaluate whether local binary pattern texture features with superpixel segmentation and classification with support vector machine can be utilized to identify immune cell infiltration in H and E-stained breast cancer samples. Guided with the pan-leukocyte CD45 marker, we annotated training and test sets from 20 primary breast cancer samples. In the training set of arbitrary sized image regions (n=1,116) a 3-fold cross-validation resulted in 98% accuracy and an area under the receiver-operating characteristic curve (AUC) of 0.98 to discriminate between immune cell -rich and - poor areas. In the test set (n=204), we achieved an accuracy of 96% and AUC of 0.99 to label cropped tissue regions correctly into immune cell -rich and -poor categories. The obtained results demonstrate strong discrimination between immune cell -rich and -poor tissue morphologies. The proposed method can provide a quantitative measurement of the degree of immune cell infiltration and applied to digitally scanned H and E-stained breast cancer samples for diagnostic purposes.

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

Science.gov (United States)

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.

Ultrastructure, inferred porosity, and gram-staining character of Methanospirillum hungatei filament termini describe a unique cell permeability for this archaeobacterium

International Nuclear Information System (INIS)

Beveridge, T.J.; Sprott, G.D.; Whippey, P.

1991-01-01

By light microscopy, Methanospirillum hungatei GP1 stains gram positive at the terminal ends of each multicellular filament and gram negative at all regions in between. This phenomenon was studied further by electron microscopy and energy-dispersive X-ray spectroscopy of Gram-stained cells, using a platinum compound to replace Gram's iodine. Crystal violet-platinum precipitates could be found only in the terminal cells of each filament, which suggested that the multilamellar plugs at the filament ends were involved with stain penetration. When sheaths were isolated by sodium dodecyl sulfate-dithiothreitol treatment, the end plugs could be ejected and their layers could be separated from one another by 0.1 M NaOH treatment. Each plug consisted of at least three individual layers; two were particulate and possessed 14.0-nm particles hexagonally arranged on their surfaces with a spacing of a = b = 18.0 nm, whereas the other was a netting of 12.5-nm holes with spacings and symmetry identical to those of the particulate layers. Optical diffraction and computer image reconstruction were used to clarify the structures of each layer in an intact plug and to provide a high-resolution image of their interdigitated structures. The holes through this composite were three to six times larger than those through the sheath. Accordingly, we propose that the terminal plugs of M. hungatei allow the access of larger solutes than does the sheath and that this is the reason why the end cells of each filament stain gram positive whereas more internal cells are gram negative. Intuitively, since the cell spacers which partition the cells from one another along the filament contain plugs identical in structure to terminal plugs, the diffusion of large solutes for these cells would be unidirectional along the filament-cell axis

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

Directory of Open Access Journals (Sweden)

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

Directory of Open Access Journals (Sweden)

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.

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

Science.gov (United States)

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.

Science.gov (United States)

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.

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

Directory of Open Access Journals (Sweden)

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.

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

Science.gov (United States)

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.

Detection of human filarial parasite Brugia malayi in dogs by histochemical staining and molecular techniques.

Science.gov (United States)

Ambily, V R; Pillai, Usha Narayana; Arun, R; Pramod, S; Jayakumar, K M

2011-09-27

Human filariasis caused by Brugia malayi is still a public health problem in many countries of Asia including India, Indonesia, Malaysia and Thailand. The World Health Organization (WHO) has targeted to eliminate filariasis by the year 2020 by Mass annual single dose Diethylcarbamazine Administration (MDA). Results of the MDA programme after the first phase was less satisfactory than expected. Malayan filariasis caused by B. malayi is endemic in the south of Thailand where domestic cat serves as the major reservoir host. There is no report about the occurrence of B. malayi in dogs. The present work was carried out to find out the incidence of microfilariasis in dogs and also to detect the presence of human filarial infection in dogs, if any. One hundred dogs above 6 months of age presented to the veterinary college Hospital, Mannuthy, Kerala, with clinical signs suggestive of microfilariasis - fever, anorexia, conjunctivitis, limb and scrotal oedema - were screened for microfilariae by wet film examination. Positive cases were subjected to Giemsa staining, histochemical staining and molecular techniques. Results of the study showed that 80% of dogs had microfilariasis; out of which 20% had sheathed microfilaria. Giemsa and histochemical staining character, PCR and sequencing confirmed it as B. malayi. High prevalence of B. malayi in dogs in this study emphasized the possible role of dogs in transmission of human filariasis. Copyright © 2011 Elsevier B.V. All rights reserved.

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.

Science.gov (United States)

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

Directory of Open Access Journals (Sweden)

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.

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.

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

Topographical distribution and morphology of NADPH-diaphorase-stained neurons in the human claustrum

Science.gov (United States)

Hinova-Palova, Dimka V.; Edelstein, Lawrence; Landzhov, Boycho; Minkov, Minko; Malinova, Lina; Hristov, Stanislav; Denaro, Frank J.; Alexandrov, Alexandar; Kiriakova, Teodora; Brainova, Ilina; Paloff, Adrian; Ovtscharoff, Wladimir

2014-01-01

We studied the topographical distribution and morphological characteristics of NADPH-diaphorase-positive neurons and fibers in the human claustrum. These neurons were seen to be heterogeneously distributed throughout the claustrum. Taking into account the size and shape of stained perikarya as well as dendritic and axonal characteristics, Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPHd)-positive neurons were categorized by diameter into three types: large, medium and small. Large neurons ranged from 25 to 35 μm in diameter and typically displayed elliptical or multipolar cell bodies. Medium neurons ranged from 20 to 25 μm in diameter and displayed multipolar, bipolar and irregular cell bodies. Small neurons ranged from 14 to 20 μm in diameter and most often displayed oval or elliptical cell bodies. Based on dendritic characteristics, these neurons were divided into spiny and aspiny subtypes. Our findings reveal two populations of NADPHd-positive neurons in the human claustrum—one comprised of large and medium cells consistent with a projection neuron phenotype, the other represented by small cells resembling the interneuron phenotype as defined by previous Golgi impregnation studies. PMID:24904317

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.

Science.gov (United States)

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.

Correlation between the proportion of stained eggs and the number of mites (Dermanyssus gallinae) monitored using a 'non-parallel board trap'.

Science.gov (United States)

Odaka, Makiko; Ogino, Kazumasa; Shikada, Michitaka; Asada, Kenichi; Kasa, Syoujirou; Inoue, Takahiro; Maeda, Ken

2017-12-01

The poultry red mite (Dermanyssus gallinae) is a serious problem for the poultry industry worldwide. However, the relationship between the mite population and the damage that they cause is still unclear. In this study, the mite population in poultry houses was examined using an established trap method, and the risk of blood-stained eggs caused by the mites was assessed. Traps were placed once a week outside the egg channels and/or on the floor in two poultry farms in Fukuoka Prefecture, Japan, from April 2012 to July 2014. The numbers of blood-stained eggs and total eggs were counted at weekly intervals. The results showed that the number of mites increased from April to May, and reached a peak around the beginning of June when the average temperature and humidity were >24°C and 70-90%, respectively. In the segmented model, the correlation between the proportion of blood-stained eggs and the number of mites or temperature was positive over a threshold. In conclusion, our established trap method is useful for monitoring mites and can be used to predict when poultry farms should be treated to prevent appearance of blood-stained eggs. © 2017 Japanese Society of Animal Science.

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.

Human herpesvirus-8 positive iatrogenic Kaposi's sarcoma in the setting of refractory ulcerative colitis.

Science.gov (United States)

Duh, Erica; Fine, Sean

2017-12-16

Although Kaposi sarcoma (KS) has been more traditionally considered an AIDS-defining illness, it may also be seen in individuals on immunosuppresive therapy. We report a case of a patient who presented to the hospital in the setting of increasingly refractory ulcerative colitis. Computed tomography scan of the abdomen was consistent with sigmoid diverticulititis and blood cultures were positive for Klebsiella. After a course of antibiotics with resolution of infection, a colonoscopy was performed to evaluate his diverticulitis and incidentally revealed a new rectal tumor. Immunohistochemistry showed the tumor was consistent with KS, with cells staining strongly positive for human herpesvirus-8. This case not only illustrates a rare case of KS found in an HIV-negative individual, but it also highlights the importance of considering an alternative diagnosis in a patient refractory to medical treatment. We discuss the management and care of an ulcerative colitis patient diagnosed with KS on immunosuppressive therapy.

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.

Anti-Jo-1 antibody-positive patients show a characteristic necrotizing perifascicular myositis.

Science.gov (United States)

Mescam-Mancini, Lénaig; Allenbach, Yves; Hervier, Baptiste; Devilliers, Hervé; Mariampillay, Kuberaka; Dubourg, Odile; Maisonobe, Thierry; Gherardi, Romain; Mezin, Paulette; Preusse, Corinna; Stenzel, Werner; Benveniste, Olivier

2015-09-01

Idiopathic inflammatory myopathies can be classified as polymyositis, dermatomyositis, immune-mediated necrotizing myopathy, sporadic inclusion body myositis or non-specific myositis. Anti-Jo-1 antibody-positive patients are assigned to either polymyositis or dermatomyositis suggesting overlapping pathological features. We aimed to determine if anti-Jo-1 antibody-positive myopathy has a specific morphological phenotype. In a series of 53 muscle biopsies of anti-Jo-1 antibody-positive patients, relevant descriptive criteria defining a characteristic morphological pattern were identified. They were tested in a second series of anti-Jo-1 antibody-positive patients and compared to 63 biopsies from patients suffering from other idiopathic inflammatory myopathies. In anti-Jo-1 antibody-positive patients, necrotic fibres, which strongly clustered in perifascicular regions, were frequently observed. Sarcolemmal complement deposition was detected specifically in perifascicular areas. Inflammation was mainly located in the perimysium and around vessels in 90.6%. Perimysial fragmentation was observed in 90% of cases. Major histocompatibility complex class I staining was diffusely positive, with a perifascicular reinforcement. Multivariate analysis showed that criteria defining perifascicular pathology: perifascicular necrosis, atrophy, and perimysial fragmentation allow the distinction of anti-Jo-1 antibody-positive patients, among patients suffering from other idiopathic inflammatory myopathies. Anti-Jo-1 antibody-positive patients displayed perifascicular necrosis, whereas dermatomyositis patients exhibited perifascicular atrophy. © The Author (2015). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

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.

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.

Strongly coupled Coulomb systems with positive dust grains: thermal and UV-induced plasmas

International Nuclear Information System (INIS)

Samarian, A.A.

2000-01-01

Full text: A plasma containing macroscopic dust particles or grains (often referred to as a dusty or colloidal or complex plasma) has the feature that grains may be charged by electron or ion flux or by photo- or thermoelectron emission. Electron emission from a grain surface produces a positive charge; capture of electrons produces the reverse effect making the dust grains negatively charged. Most dusty plasma research is concerned with the ordered dust structures (so-called 'plasma crystal') in glow discharges. The dust grains in these experiments were found to carry a negative charge due to the higher mobility of electrons as compared to ions in the discharge plasma. In recent years, in parallel with the study of the properties of plasma crystals under discharge conditions, attempts to obtain a structure from positively charged dust grains have been made, and structure formation processes for various charging mechanisms, particularly thermoelectron emission and photoemission, have been investigated. In this paper we review the essential features of strongly coupled plasmas with positive dust grains. An ordered structure of CeO 2 grains has been experimentally observed in a combustion products jet. The grains were charged positively and suspended in the plasma flow. Their charge is about 10 3 a and the calculated value of a Coulomb coupling parameter Γ is >10, corresponding to a plasma liquid. The ordered structures of Al 2 O 3 dust grains in propellant combustion products plasma have been observed for the first time. These structures were found in the sheath boundary of condensation region. The obtained data let us estimate the value of parameter Γ =3-40, corresponding to the plasma liquid state. The possibility is studied of the formation of ordered dust grain structures in thermal plasma. The range of the required values of the coupling parameter Γ is calculated using the results of diagnostic measurements carried out in thermal plasma with grains of

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.

Superselective intra-arterial hepatic injection of indocyanine green (ICG) for fluorescence image-guided segmental positive staining: experimental proof of the concept.

Science.gov (United States)

Diana, Michele; Liu, Yu-Yin; Pop, Raoul; Kong, Seong-Ho; Legnèr, Andras; Beaujeux, Remy; Pessaux, Patrick; Soler, Luc; Mutter, Didier; Dallemagne, Bernard; Marescaux, Jacques

2017-03-01

Intraoperative liver segmentation can be obtained by means of percutaneous intra-portal injection of a fluorophore and illumination with a near-infrared light source. However, the percutaneous approach is challenging in the minimally invasive setting. We aimed to evaluate the feasibility of fluorescence liver segmentation by superselective intra-hepatic arterial injection of indocyanine green (ICG). Eight pigs (mean weight: 26.01 ± 5.21 kg) were involved. Procedures were performed in a hybrid experimental operative suite equipped with the Artis Zeego ® , multiaxis robotic angiography system. A pneumoperitoneum was established and four laparoscopic ports were introduced. The celiac trunk was catheterized, and a microcatheter was advanced into different segmental hepatic artery branches. A near-infrared laparoscope (D-Light P, Karl Storz) was used to detect the fluorescent signal. To assess the correspondence between arterial-based fluorescence demarcation and liver volume, metallic markers were placed along the fluorescent border, followed by a 3D CT-scanning, after injecting intra-arterial radiological contrast (n = 3). To assess the correspondence between arterial and portal supplies, percutaneous intra-portal angiography and intra-arterial angiography were performed simultaneously (n = 1). Bright fluorescence signal enhancing the demarcation of target segments was obtained from 0.1 mg/mL, in matter of seconds. Correspondence between the volume of hepatic segments and arterial territories was confirmed by CT angiography. Higher background fluorescence noise was found after positive staining by intra-portal ICG injection, due to parenchymal accumulation and porto-systemic shunting. Intra-hepatic arterial ICG injection, rapidly highlights hepatic target segment borders, with a better signal-to-background ratio as compared to portal vein injection, in the experimental setting.

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.

Comparison of different stains in imprint cytology with the conventional diagnostic tools in detection of Helicobacter pylori infection

Directory of Open Access Journals (Sweden)

Shashikant Adlekha

2015-01-01

Full Text Available Background: Helicobacter pylori (H. pylori infection is one of the most common causes of gastrointestinal morbidity and mortality worldwide. Early diagnosis of infection is imperative for the cure of distressing symptoms and prevention of complications. Objective: The objective of the study is to evaluate imprint cytology of gastric biopsy specimens as a rapid and cost-effective diagnostic tool and its comparison with conventional diagnostic tests. Materials and Methods: Antral biopsies were collected from 130 patients and evaluated for H. pylori infection by imprint cytology and histopathological examination by different stains and rapid urease test (RUT. Histopathological features of gastric biopsy specimens were also assessed. Results: A total of 118 patients showed H. pylori infection by two or more methods. Giemsa histology showed highest sensitivity, specificity, positive predictive value, negative predictive value and Youden′s Index (YI. Among imprint cytology stain methods, toluidine blue showed highest sensitivity and highest YI was obtained for PAP stain. Conclusion: Helicobacter pylori infection is associated with gastric mucosa changes like chronic active gastritis, atrophy, intestinal metaplasia, ulceration and carcinoma. Imprint cytology has high sensitivity and comparable predictive values to conventional diagnostic tools-histopathological examination and RUT in the detection of H. pylori infection.

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.

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.

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.

[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.

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

[Clinical usefulness of urine-formed elements' information obtained from bacteria detection by flow cytometry method that uses nucleic acid staining].

Science.gov (United States)

Nakagawa, Hiroko; Yuno, Tomoji; Itho, Kiichi

2009-03-01

Recently, specific detection method for Bacteria, by flow cytometry method using nucleic acid staining, was developed as a function of automated urine formed elements analyzer for routine urine testing. Here, we performed a basic study on this bacteria analysis method. In addition, we also have a comparison among urine sediment analysis, urine Gram staining and urine quantitative cultivation, the conventional methods performed up to now. As a result, the bacteria analysis with flow cytometry method that uses nucleic acid staining was excellent in reproducibility, and higher sensitivity compared with microscopic urinary sediment analysis. Based on the ROC curve analysis, which settled urine culture method as standard, cut-off level of 120/microL was defined and its sensitivity = 85.7%, specificity = 88.2%. In the analysis of scattergram, accompanied with urine culture method, among 90% of rod positive samples, 80% of dots were appeared in the area of 30 degrees from axis X. In addition, one case even indicated that analysis of bacteria by flow cytometry and scattergram of time series analysis might be helpful to trace the progress of causative bacteria therefore the information supposed to be clinically significant. Reporting bacteria information with nucleic acid staining flow cytometry method is expected to contribute to a rapid diagnostics and treatment of urinary tract infections. Besides, the contribution to screening examination of microbiology and clinical chemistry, will deliver a more efficient solution to urine analysis.

Gram-stain plus MALDI-TOF MS (Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for a rapid diagnosis of urinary tract infection.

Directory of Open Access Journals (Sweden)

Almudena Burillo

Full Text Available Microbiological confirmation of a urinary tract infection (UTI takes 24-48 h. In the meantime, patients are usually given empirical antibiotics, sometimes inappropriately. We assessed the feasibility of sequentially performing a Gram stain and MALDI-TOF MS mass spectrometry (MS on urine samples to anticipate clinically useful information. In May-June 2012, we randomly selected 1000 urine samples from patients with suspected UTI. All were Gram stained and those yielding bacteria of a single morphotype were processed for MALDI-TOF MS. Our sequential algorithm was correlated with the standard semiquantitative urine culture result as follows: Match, the information provided was anticipative of culture result; Minor error, the information provided was partially anticipative of culture result; Major error, the information provided was incorrect, potentially leading to inappropriate changes in antimicrobial therapy. A positive culture was obtained in 242/1000 samples. The Gram stain revealed a single morphotype in 207 samples, which were subjected to MALDI-TOF MS. The diagnostic performance of the Gram stain was: sensitivity (Se 81.3%, specificity (Sp 93.2%, positive predictive value (PPV 81.3%, negative predictive value (NPV 93.2%, positive likelihood ratio (+LR 11.91, negative likelihood ratio (-LR 0.20 and accuracy 90.0% while that of MALDI-TOF MS was: Se 79.2%, Sp 73.5, +LR 2.99, -LR 0.28 and accuracy 78.3%. The use of both techniques provided information anticipative of the culture result in 82.7% of cases, information with minor errors in 13.4% and information with major errors in 3.9%. Results were available within 1 h. Our serial algorithm provided information that was consistent or showed minor errors for 96.1% of urine samples from patients with suspected UTI. The clinical impacts of this rapid UTI diagnosis strategy need to be assessed through indicators of adequacy of treatment such as a reduced time to appropriate empirical treatment or

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

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.

Fascin and EMMPRIN expression in primary mucinous tumors of ovary: a tissue microarray study.

Science.gov (United States)

Alici, Omer; Kefeli, Mehmet; Yildiz, Levent; Baris, Sancar; Karagoz, Filiz; Kandemir, Bedri

2014-12-01

The aim of this study was to compare the expressions of fascin and EMMPRIN in primary malignant, borderline and benign mucinous ovarian tumors, and to investigate the relationship of these markers with tumor progression and their applicability to differential diagnosis. An immunohistochemical study was performed for fascin and EMMPRIN using the tissue microarray technique. Eighty-one cases were included in the study; there were 37 benign, 25 borderline and 19 malignant primary mucinous ovarian tumors. For each case, a total staining score was determined, consisting of scores for extent of staining and intensity of staining. The cases were allocated to negative, weakly positive and strongly positive staining categories, according to the total staining score. Both of the markers were significantly negative in benign tumors as compared with borderline and malignant tumors. There was no significant difference between borderline and malignant groups for both markers. Sixty-eight percent of malignant tumors were stained positive by fascin, while this rate was 40% for borderline mucinous tumors. All malignant tumors were strongly stained positive for EMMPRIN, while this rate was 92% for borderline mucinous tumors. The rest of the cases stained weakly positive. No significant difference in staining score was found between fascin and EMMPRIN expression. In ovarian primary mucinous tumors, fascin and EMMPRIN may play an important role in tumor progression from benign tumor to carcinoma. In that context, EMMPRIN and fascin expression may have potential application in the differential diagnosis of some diagnostically problematic mucinous ovarian tumors. However, the differential diagnostic applicability of EMMPRIN appears to be more limited than that of fascin due to its wide spectrum of staining in mucinous ovarian tumors. Copyright © 2014 Elsevier GmbH. All rights reserved.

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.

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

Diffuse reflectance FTIR of stains on grit blasted metals

Energy Technology Data Exchange (ETDEWEB)

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

1997-08-09

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

Visualisation of plastid outgrowths in potato (Solanum tuberosum L.) tubers by carboxyfluorescein diacetate staining.

Science.gov (United States)

Borucki, Wojciech; Bederska, Magdalena; Sujkowska-Rybkowska, Marzena

2015-05-01

We describe two types of plastid outgrowths visualised in potato tubers after carboxyfluorescein diacetate staining. Probable esterase activity of the outgrowths has been demonstrated for the first time ever. Plastid outgrowths were observed in the phelloderm and storage parenchyma cells of red potato (S. tuberosum L. cv. Rosalinde) tubers after administration of carboxyfluorescein diacetate stain. Endogenous esterases cleaved off acetic groups to release membrane-unpermeable green fluorescing carboxyfluorescein which accumulated differentially in particular cell compartments. The intensive green fluorescence of carboxyfluorescein exhibited highly branched stromules (stroma-filled plastid tubular projections of the plastid envelope) and allowed distinguishing them within cytoplasmic strands of the phelloderm cells. Stromules (1) were directed towards the nucleus or (2) penetrated the whole cells through the cytoplasmic bands of highly vacuolated phelloderm cells. Those directed towards the nucleus were flattened and adhered to the nuclear envelope. Stromule-like interconnections between two parts of the same plastids (isthmuses) were also observed. We also documented the formation of another type of the stroma-filled plastid outgrowths, referred to here as protrusions, which differed from previously defined stromules in both morphology and esterase activity. Unlike stromules, the protrusions were found to be associated with developmental processes leading to starch accumulation in the storage parenchyma cells. These results strongly suggest that stromules and protrusions exhibit esterase activity. This has been demonstrated for the first time. Morphological and biochemical features as well as possible functions of stromules and protrusions are discussed below.

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.

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.

Automated quantification of proliferation with automated hot-spot selection in phosphohistone H3/MART1 dual-stained stage I/II melanoma.

Science.gov (United States)

Nielsen, Patricia Switten; Riber-Hansen, Rikke; Schmidt, Henrik; Steiniche, Torben

2016-04-09

Staging of melanoma includes quantification of a proliferation index, i.e., presumed melanocytic mitoses of H&E stains are counted manually in hot spots. Yet, its reproducibility and prognostic impact increases by immunohistochemical dual staining for phosphohistone H3 (PHH3) and MART1, which also may enable fully automated quantification by image analysis. To ensure manageable workloads and repeatable measurements in modern pathology, the study aimed to present an automated quantification of proliferation with automated hot-spot selection in PHH3/MART1-stained melanomas. Formalin-fixed, paraffin-embedded tissue from 153 consecutive stage I/II melanoma patients was immunohistochemically dual-stained for PHH3 and MART1. Whole slide images were captured, and the number of PHH3/MART1-positive cells was manually and automatically counted in the global tumor area and in a manually and automatically selected hot spot, i.e., a fixed 1-mm(2) square. Bland-Altman plots and hypothesis tests compared manual and automated procedures, and the Cox proportional hazards model established their prognostic impact. The mean difference between manual and automated global counts was 2.9 cells/mm(2) (P = 0.0071) and 0.23 cells per hot spot (P = 0.96) for automated counts in manually and automatically selected hot spots. In 77 % of cases, manual and automated hot spots overlapped. Fully manual hot-spot counts yielded the highest prognostic performance with an adjusted hazard ratio of 5.5 (95 % CI, 1.3-24, P = 0.024) as opposed to 1.3 (95 % CI, 0.61-2.9, P = 0.47) for automated counts with automated hot spots. The automated index and automated hot-spot selection were highly correlated to their manual counterpart, but altogether their prognostic impact was noticeably reduced. Because correct recognition of only one PHH3/MART1-positive cell seems important, extremely high sensitivity and specificity of the algorithm is required for prognostic purposes. Thus, automated

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

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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

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.

Comparison between morphological and staining characteristics of live and dead eggs of Schistosoma mansoni

Directory of Open Access Journals (Sweden)

AK Sarvel

2006-10-01

Full Text Available Schistosoma mansoni eggs are classified, according to morphological characteristics, as follows: viable mature and immature eggs; dead mature and immature eggs, shells and granulomas. The scope of this study was to compare the staining characteristics of different morphological types of eggs in the presence of fluorescent labels and vital dyes, aiming at differentiating live and dead eggs. The eggs were obtained from the intestines of infected mice, and put into saline 0.85%. The fluorescent labels were Hoechst 33258 and Acridine Orange + Ethidium Bromide and vital dyes (Trypan Blue 0.4% and Neutral Red 1%. When labelled with the probe Hoechst 33258, some immature eggs, morphologically considered viable, presented fluorescence (a staining characteristic detected only in dead eggs; mature eggs did not present fluorescence, and the other types of dead eggs, morphologically defined, showed fluorescence. As far as Acridine Orange + Ethidium Bromide are concerned, either the eggs considered to be live, or the dead ones, presented staining with green color, and only the hatched and motionless miracidium was stained with an orange color. Trypan Blue was not able to stain the eggs, considered to be dead but only dead miracidia which had emerged out of the shell. Neutral Red stained both live and dead eggs. Only the fluorescent Hoechst 33258 can be considered a useful tool for differentiation between dead and live eggs.

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.

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

p16/ki-67 dual-stain cytology in the triage of ASCUS and LSIL papanicolaou cytology: results from the European equivocal or mildly abnormal Papanicolaou cytology study.

Science.gov (United States)

Schmidt, Dietmar; Bergeron, Christine; Denton, Karin J; Ridder, Ruediger

2011-06-25

The objective of this study was to analyze the diagnostic performance of a newly established immunocytochemical dual-stain protocol, which simultaneously detects p16(INK4a) and Ki-67 expression in cervical cytology samples, for identifying high-grade cervical intraepithelial neoplasia (CIN2+) in women with Papanicolaou (Pap) cytology results categorized as atypical squamous cells of undetermined significance (ASCUS) or low-grade squamous intraepithelial lesions (LSIL). Residual liquid-based cytology material from 776 retrospectively collected ASCUS/LSIL cases that were available from a recent study evaluating p16 cytology and HPV testing were subjected to p16/Ki-67 dual staining. The presence of 1 or more double-immunoreactive cell(s) was regarded as a positive test outcome, irrespective of morphology. Test results were correlated to histology follow-up. Sensitivity of p16/Ki-67 dual-stain cytology for biopsy-confirmed CIN2+ was 92.2% (ASCUS) and 94.2% (LSIL), while specificity rates were 80.6% (ASCUS) and 68.0% (LSIL), respectively. Similar sensitivity/specificity profiles were found for both age groups of women aged aged ≥30 years. Dual-stain cytology showed comparable sensitivity, but significantly higher specificity, when compared with human papillomavirus (HPV) testing. The results of this study show that p16/Ki-67 dual-stain cytology provided a high sensitivity for the detection of underlying CIN2+ in women with ASCUS or LSIL Pap cytology results, comparable to the rates previously reported for HPV testing and p16 single-stain cytology. However, the specificity of this morphology-independent interpretation of p16/Ki-67 dual-stain cytology testing was further improved compared with the earlier p16 single-stain cytology approach, which required morphology interpretation, and it is significantly higher when compared with HPV testing. Copyright © 2011 American Cancer Society.

Is hematoxylin-eosin staining in rectal mucosal and submucosal biopsies still useful for the diagnosis of Hirschsprung disease?

Science.gov (United States)

Serafini, Suellen; Santos, Maria Mercês; Aoun Tannuri, Ana Cristina; Zerbini, Maria Claudia Nogueira; de Mendonça Coelho, Maria Cecília; de Oliveira Gonçalves, Josiane; Tannuri, Uenis

2017-12-06

Hematoxylin-eosin (HE) staining of a full-thickness rectal wall fragment is classically used for the diagnosis of Hirschsprung disease (HD). However, this technique requires large fragments for a better diagnosis. Additionally, the histochemical and immunohistochemical methods of staining small fragments of rectal mucosal and submucosal biopsies are not available in all centers. Therefore, the possibility of diagnosing HD through HE staining in these biopsies could be a valuable alternative for centers that do not have more specific techniques. The objectives of the current investigation were to evaluate the concordance of the results obtained by HE staining and the calretinin method with acetylcholinesterase (AChE) activity in fragments of mucosa and submucosa in the diagnosis of HD. For this study, 50 cases from our laboratory were selected. The tissue material was embedded in paraffin. Sixty levels of each fragment were utilized for HE, and the other 3 levels were used for calretinin. These slides were analyzed under the microscope, photographed and classified as either positive for HD when no ganglion cells were found with nerve trunks present or as negative when ganglion cells were found. The results from reading the slides were compared with those of AChE. Of the 50 cases evaluated by the HE technique, only 5 contradicted the diagnosis based on AChE, with a Kappa value of 0.800 and an accuracy of 90%. In the comparison between calretinin and AChE, 8 cases were discordant, with a Kappa value of 0.676 and an accuracy of 84%. The concordance of results from AChE and HE methods was satisfactory, allowing for the potential use of the HE method for fragments of mucosa and submucosa as a valid alternative in the diagnosis of HD. The immunohistochemical technique of calretinin did not show good agreement with the AChE activity in our study.

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.

Ultrastructural demonstration of a glycoproteinic surface coat in allergenic pollen grains by combined cetylpyridinium chloride precipitation and silver proteinate staining.

Science.gov (United States)

Grote, M; Fromme, H G

1984-01-01

In allergenic birch pollen grains, highly watersoluble surface substances were precipitated by the cationic detergent cetylpyridinium chloride (CPC) during aqueous fixation. After processing the pollen for electron microscopy, ultrathin sections of pollen grains were subjected to the periodic acid - thiocarbohydrazide - silver proteinate (PA-TCH-SP) procedure according to Thiery (1967) for the detection of vicinal glycol groups. It was found that the material precipitated by CPC on the surface and within the exine cavities of the pollen wall strongly reacted with the PA-TCH-SP reagent thus indicating the presence of polysaccharides on the surface of birch pollen grains. In samples which had not been treated with the cationic detergent, PA-TCH-SP reactivity was reduced to thin linings on the surface and within the exine cavities. In both cases the exine proper did not stain whereas the intine showed moderate staining. Within the aperture region of the intine, PA-TCH-SP reactivity is preferably associated with fibrillar or reticular structures. The results are discussed with special reference to biochemical findings on allergenic birch pollen proteins.

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.

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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

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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

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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

The development of fluorescence turn-on probe for Al(III) sensing and live cell nucleus-nucleoli staining

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Saini, Anoop Kumar; Sharma, Vinay; Mathur, Pradeep; Shaikh, Mobin M.

2016-10-01

The morphology of nucleus and nucleolus is powerful indicator of physiological and pathological conditions. The specific staining of nucleolus recently gained much attention due to the limited and expensive availability of the only existing stain “SYTO RNA-Select”. Here, a new multifunctional salen type ligand (L1) and its Al3+ complex (1) are designed and synthesized. L1 acts as a chemosensor for Al3+ whereas 1 demonstrates specific staining of nucleus as well as nucleoli. The binding of 1 with nucleic acid is probed by DNase and RNase digestion in stained cells. 1 shows an excellent photostability, which is a limitation for existing nucleus stains during long term observations. 1 is assumed to be a potential candidate as an alternative to expensive commercial dyes for nucleus and nucleoli staining.

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.

Science.gov (United States)

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

Exact solutions of strong gravity in generalized metrics

International Nuclear Information System (INIS)

Hojman, R.; Smailagic, A.

1981-05-01

We consider classical solutions for the strong gravity theory of Salam and Strathdee in a wider class of metrics with positive, zero and negative curvature. It turns out that such solutions exist and their relevance for quark confinement is explored. Only metrics with positive curvature (spherical symmetry) give a confining potential in a simple picture of the scalar hadron. This supports the idea of describing the hadron as a closed microuniverse of the strong metric. (author)

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.

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.

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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.

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.

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.

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

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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.

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.

Prevalence of mabDAS-1 positivity in biopsy specimens from the esophagogastric junction.

Science.gov (United States)

Rogge-Wolf, Claudia; Seldenrijk, Cornelis A; Das, Kiron M; Timmer, Robin; Breumelhof, Ronald; Smout, André J P M; Amenta, Peter S; Griffel, Louis H

2002-12-01

Intestinal metaplasia (IM) is a precursor for malignancies at the esophagogastric junction. A monoclonal antibody, mAbDAS-1, can probably identify cellular characteristics of IM before the appearance of goblet cells. The aim of this study was to examine the prevalence of mAbDAS-1 positivity in biopsies from the squamocolumnar junction (SCJ) and to correlate this positivity with the presence of IM and clinical findings. In 559 patients, reflux symptoms were scored, and the presence of reflux esophagitis and hiatus hernia was evaluated during endoscopy. Two biopsy specimens were obtained from the SCJ. In a subset of patients (n = 99), biopsies from the endoscopically defined cardiac region (2 cm distal to proximal margin of gastric folds) were available. Biopsy specimens were stained with hematoxylin and eosin, Alcian Blue, modified Giemsa, and mAbDAS-1. mAbDAS-1 positivity was observed in the SCJ biopsies of 201 of 486 (41.4%) patients without IM and in 64 of 73 (87.7%) patients with IM. Patients without IM but with antibody positivity showed similar histological characteristics as patients with IM at the SCJ. Biopsies of 123 of 559 patients (22%) revealed a columnar-cuboidal epithelium, which was found to be mAbDAS-1 positive in 64.2% (77 of 123). Tissue specimens from the cardiac region without IM stained positive in 14.2% (13 of 91), 12 of those also stained at the SCJ. In patients without IM, a high prevalence of mAbDAS-1 positivity was observed. Biopsies of these patients showed similar histological characteristics as patients with IM. Although not all patients exhibiting this reactivity may develop IM, mAbDAS-1 reactivity may help in the understanding of the histogenesis of IM at the SCJ.

Staining Against Phospho-H2AX (gamma-H2AX) as a Marker for DNA Damage and Genomic Instability in Cancer Tissues and Cells

NARCIS (Netherlands)

Nagelkerke, A.P.; Span, P.N.

2016-01-01

Phospho-H2AX or gamma-H2AX- is a marker of DNA double-stranded breaks and can therefore be used to monitor DNA repair after, for example, irradiation. In addition, positive staining for phospho-H2AX may indicate genomic instability and telomere dysfunction in tumour cells and tissues. Here, we

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.

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)

Weathering patinas on the medieval (S. XIV) stained glass windows of the Pedralbes Monastery (Barcelona, Spain).

Science.gov (United States)

Aulinas, Meritxell; Garcia-Valles, Maite; Gimeno, Domingo; Fernandez-Turiel, Jose Luis; Ruggieri, Flavia; Pugès, Montserrat

2009-06-01

contrast, weddellite is limited to the absence of thenardite and whewellite and to the external parts of the stained glass. Quartz is present in all the patinas independent of their location and color. Calcite also occurs in many samples. It appears in both color-type patinas and, in some cases, is associated to the presence of weddellite but not to whewellite and/or thenardite. Glass composition together with environmental conditions and location of the patinas (internal or external parts of the stained glass window), as well as the provenance of the glass within the monastery, are the main factors that define the development of the new mineral phases. Moreover, the action of microorganisms, when present, can also strongly influence the development of some mineral phases. For example, the formation of calcite in the external parts of the stained glass (associated with the presence of oxalates) is related to the action of microorganisms. When calcite is formed in the internal parts of the glass and it is not associated with the presence of Ca-oxalates, an inorganic origin can be invoked. The presence of weddellite requires a very humid microenvironment with very little exposure to sunlight. In fact, this mineral phase has only been observed in the external parts of some glass located in the humid and shady side of the monastery. Whewellite (which only appears in the internal parts) needs a low degree of relative humidity. It has been observed that sulfur precipitating in basically one mineral phase (thenardite or gypsum) depends on the microenvironmental conditions of the moment and the glass composition. When thenardite occurs, it can be maintained that the original glass is of Na composition. The occurrence of quartz in all samples is interpreted as being due to the deposition of atmospheric particulate matter. The color of the patinas can be originated by different processes (presence of carotenes, organic pigmentation, atmospheric contamination, etc.). In the case of

Time Course of Detection of Human Male DNA from Stained Blood Sample on Various Surfaces by Loop Mediated Isothermal Amplification and Polymerase Chain Reaction

Directory of Open Access Journals (Sweden)

Panan Kanchanaphum

2018-01-01

Full Text Available This study explores determining the sex of humans from blood stains taken from different surfaces and compares the time course of detection with the conventional PCR, Conventional Loop Mediated Isothermal Amplification (LAMP, and LAMP-Lateral Flow Dipstick (LFD. For the DNA templates, 7 male and 7 female blood stained samples were extracted and added to LAMP and PCR reaction solution to amplify the SRY gene. The DNA samples were extracted from the following blood stained materials: cloth, wood, clay, and tile. Then, the samples were stored at room temperature for 1, 7, 30, and 60 day(s. After the DNA amplification, the gel electrophoresis process was applied to detect LAMP product. The LFD was combined with the LAMP to detect LAMP product on the male cloth samples. For the male samples, the time course of detection on the first and seventh days indicated positive for both LAMP and PCR products on all the surfaces while no DNA amplification was found on any of the female samples. On day 30, positive LAMP product was still found on all the male samples. However, it had faded on the tiles. Moreover, all the male samples, which had tested positive for PCR product, were blurred and unclear. On day 60, LAMP product was still found on all the male samples. Conversely, the PCR method resulted in no bands showing for any of the male samples. However, the LAMP-LFD method detected product on all the male samples of cloth. The results show that the LAMP is an effective, practical, and reliable molecular-biological method. Moreover, the LFD can increase the efficiency and sensitivity of the LAMP, making it more suitable for field studies because gel electrophoresis apparatus is not required.

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.

Strong monotonicity in mixed-state entanglement manipulation

International Nuclear Information System (INIS)

Ishizaka, Satoshi

2006-01-01

A strong entanglement monotone, which never increases under local operations and classical communications (LOCC), restricts quantum entanglement manipulation more strongly than the usual monotone since the usual one does not increase on average under LOCC. We propose strong monotones in mixed-state entanglement manipulation under LOCC. These are related to the decomposability and one-positivity of an operator constructed from a quantum state, and reveal geometrical characteristics of entangled states. These are lower bounded by the negativity or generalized robustness of entanglement

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

Defensive marketing: how a strong incumbent can protect its position.

Science.gov (United States)

Roberts, John H

2005-11-01

There has been a lot of research on marketing as an offensive tactic-how it can help companies successfully launch new products, enter new markets, or gain share with existing products in their current markets. But for nearly every new product launch, market entrant, or industry upstart grabbing market share, there is an incumbent that must defend its position. And there has been little research on how these defenders can use marketing to preemptively respond to new or anticipated threats. John H. Roberts outlines four basic types of defensive marketing strategies: positive, inertial, parity, and retarding. With the first two, you establish and communicate your points of superiority relative to the new entrant; with the second two, you establish and communicate strategic points of comparability with your rival. Before choosing a strategy, you need to assess the weapons you have available to protect your market position-your brand identity, the products and services that support that identity, and your means of communicating it. Then assess your customers' value to you and their vulnerability to being poached by rivals. The author explains how Australian telecommunications company Telstra, facing deregulation, used a combination of the four strategies (plus the author's customer response model) to fend off market newcomer Optus. Telstra was prepared, for instance, to reach deep into its pockets and engage in a price war. But the customer response model indicated that a parity strategy-in which Telstra would offer lower rates on some routes and at certain times of day, even though its prices, on average, were higher than its rival's-was more likely to prevent consumers from switching. Ultimately, Telstra was able to retain several points of market share it otherwise would have lost. The strategies described here, though specific to Telstra's situation, offer lessons for any company facing new and potentially damaging competition.

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

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.

TEM validation of immunohistochemical staining prior to assessment of tumour angiogenesis by computerised image analysis

International Nuclear Information System (INIS)

Killingsworth, M.C.

2002-01-01

could be evaluated using computerised image analysis. Vessel area, aspect ratio, roundness and dendritic pattern were selected as parameters for measurement. ii) Assessment of other cell types associated with growing new vessels. Initial attempts at automated measurement of angiogenic parameters were made difficult by high numbers of CD34 positive cell processes surrounding tumour glands in association with blood vessels. EM examination revealed fibroblasts, lymphocytes and macrophages in close contact with developing endothelial sprouts. Fibroblasts in particular displayed slender elongated cell processes that could be confused with endothelial sprouts. This raised the question as to whether anti-CD34 staining was cross-reacting with these other cell types, iii) Immunocytochemical assessment of anti-CD34 specificity. Immunoelectron microscopic studies to check for anti-CD34 cross-reactive staining of fibroblasts associated with growing new vessels will now be carried out. Copyright (2002) Australian Society for Electron Microscopy Inc

3D imaging of hematoxylin and eosin stained thick tissues with a sub-femtoliter resolution by using Cr:forsterite-laser-based nonlinear microscopy (Conference Presentation)

Science.gov (United States)

Kao, Chien-Ting; Wei, Ming-Liang; Liao, Yi-Hua; Sun, Chi-Kuang

2017-02-01

Intraoperative assessment of excision tissues during cancer surgery is clinically important. The assessment is used to be guided by the examination for residual tumor with frozen pathology, while it is time consuming for preparation and is with low accuracy for diagnosis. Recently, reflection confocal microscopy (RCM) and nonlinear microscopy (NLM) were demonstrated to be promising methods for surgical border assessment. Intraoperative RCM imaging may enable detection of residual tumor directly on skin cancers patients during Mohs surgery. The assessment of benign and malignant breast pathologies in fresh surgical specimens was demonstrated by NLM. Without using hematoxylin and eosin (H and E) that are common dyes for histopathological diagnosis, RCM was proposed to image in vivo by using aluminum chloride for nuclear contrast on surgical wounds directly, while NLM was proposed to detect two photon fluorescence nuclear contrast from acrdine orange staining. In this paper, we propose and demonstrate 3D imaging of H and E stained thick tissues with a sub-femtoliter resolution by using Cr:forsterite-laser-based NLM. With a 1260 nm femtosecond Cr:forsterite laser as the excitation source, the hematoxylin will strongly enhance the third-harmonic generation (THG) signals, while eosin will illuminate strong fluorescence under three photon absorption. Compared with previous works, the 1260 nm excitation light provide high penetration and low photodamage to the exercised tissues so that the possibility to perform other follow-up examination will be preserved. The THG and three-photon process provides high nonlinearity so that the super resolution in 3D is now possible. The staining and the contrast of the imaging is also fully compatible with the current clinical standard on frozen pathology thus facilitate the rapid intraoperative assessment of excision tissues. This work is sponsored by National Health Research Institutes and supported by National Taiwan University

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.

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.

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

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.

Cystic neutrophilic granulomatous mastitis: an underappreciated pattern strongly associated with gram-positive bacilli.

Science.gov (United States)

Renshaw, Andrew A; Derhagopian, Robert P; Gould, Edwin W

2011-09-01

Although granulomatous lobular mastitis is associated with gram-positive bacilli such as Corynebacterium, this association is not well known. We report 3 cases of mastitis caused by gram-positive bacilli. All 3 abscesses were suppurative with distinct enlarged cystic spaces in which rare gram-positive bacilli were identified. Two cases were also granulomatous. Cultures in all 3 cases were negative. All 3 patients recovered after biopsy and tetracycline-based therapy. Infection in the breast by gram-positive bacilli is associated with a distinct histologic pattern, including cystic spaces in the setting of neutrophilic/granulomatous inflammation that can be recognized and should prompt careful search for the organism within enlarged vacuoles.

[Fungal biomass estimation in soils from southwestern Buenos Aires province (Argentina) using calcofluor white stain].

Science.gov (United States)

Vázquez, María B; Amodeo, Martín R; Bianchinotti, María V

Soil microorganisms are vital for ecosystem functioning because of the role they play in soil nutrient cycling. Agricultural practices and the intensification of land use have a negative effect on microbial activities and fungal biomass has been widely used as an indicator of soil health. The aim of this study was to analyze fungal biomass in soils from southwestern Buenos Aires province using direct fluorescent staining and to contribute to its use as an indicator of environmental changes in the ecosystem as well as to define its sensitivity to weather conditions. Soil samples were collected during two consecutive years. Soil smears were prepared and stained with two different concentrations of calcofluor, and the fungal biomass was estimated under an epifluorescence microscope. Soil fungal biomass varied between 2.23 and 26.89μg fungal C/g soil, being these values in the range expected for the studied soil type. The fungal biomass was positively related to temperature and precipitations. The methodology used was reliable, standardized and sensitive to weather conditions. The results of this study contribute information to evaluate fungal biomass in different soil types and support its use as an indicator of soil health for analyzing the impact of different agricultural practices. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Starting off on the right foot: strong right-footers respond faster with the right foot to positive words and with the left foot to negative words.

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de la Vega, Irmgard; Graebe, Julia; Härtner, Leonie; Dudschig, Carolin; Kaup, Barbara

2015-01-01

Recent studies have provided evidence for an association between valence and left/right modulated by handedness, which is predicted by the body-specificity hypothesis (Casasanto, 2009) and also reflected in response times. We investigated whether such a response facilitation can also be observed with foot responses. Right-footed participants classified positive and negative words according to their valence by pressing a key with their left or right foot. A significant interaction between valence and foot only emerged in the by-items analysis. However, when dividing participants into two groups depending on the strength of their footedness, an interaction between valence and left/right was observed for strong right-footers, who responded faster with the right foot to positive words, and with the left foot to negative words. No interaction emerged for weak right-footers. The results strongly support the assumption that fluency lies at the core of the association between valence and left/right.

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.

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

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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

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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.

Prolyl isomerase Pin1 is highly expressed in Her2-positive breast cancer and regulates erbB2 protein stability

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Lu Kun

2008-12-01

Full Text Available Abstract Overexpression of HER-2/Neu occurs in about 25–30% of breast cancer patients and is indicative of poor prognosis. While Her2/Neu overexpression is primarily a result of erbB2 amplification, it has recently been recognized that erbB2 levels are also regulated on the protein level. However, factors that regulate Her2/Neu protein stability are less well understood. The prolyl isomerase Pin1 catalyzes the isomerization of specific pSer/Thr-Pro motifs that have been phosphorylated in response to mitogenic signaling. We have previously reported that Pin1-catalyzed post-phosphorylational modification of signal transduction modulates the oncogenic pathways downstream from c-neu. The goal of this study was to examine the expression of prolyl isomerase Pin1 in human Her2+ breast cancer, and to study if Pin1 affects the expression of Her2/Neu itself. Methods Immunohistochemistry for Her2 and Pin1 were performed on two hundred twenty-three human breast cancers, with 59% of the specimen from primary cancers and 41% from metastatic sites. Pin1 inhibition was achieved using siRNA in Her2+ breast cancer cell lines, and its effects were studied using cell viability assays, immunoblotting and immunofluorescence. Results Sixty-four samples (28.7% stained positive for Her2 (IHC 3+, and 54% (122/223 of all breast cancers stained positive for Pin1. Of the Her2-positive cancers 40 (62.5% were also Pin1-positive, based on strong nuclear or nuclear and cytoplasmic staining. Inhibition of Pin1 via RNAi resulted in significant suppression of Her2-positive tumor cell growth in BT474, SKBR3 and AU565 cells. Pin1 inhibition greatly increased the sensitivity of Her2-positive breast cancer cells to the mTOR inhibitor Rapamycin, while it did not increase their sensitivity to Trastuzumab, suggesting that Pin1 might act on Her2 signaling. We found that Pin1 interacted with the protein complex that contains ubiquitinated erbB2 and that Pin1 inhibition accelerated erbB2

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

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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

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.

Karyotype characterization of Trigona fulviventris Guérin, 1835 (Hymenoptera, Meliponini by C banding and fluorochrome staining: report of a new chromosome number in the genus

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Domingues Alayne Magalhães Trindade

2005-01-01

Full Text Available Although many species of the genus Trigona have been taxonomically described, cytogenetic studies of these species are still rare. The aim of the present study was to obtain cytogenetic data by conventional staining, C banding and fluorochrome staining for the karyotype characterization of the species Trigona fulviventris. Cytogenetic analysis revealed that this species possesses a diploid chromosome number of 2n = 32, different from most other species of this genus studied so far. This variation was probably due to the centric fusion in a higher numbered ancestral karyotype, this fusion producing the large metacentric chromosome pair and the lower chromosome number observed in Trigona fulviventris. Heterochromatin was detected in the pericentromeric region of the first chromosome pair and in one of the arms of the remaining pairs. Base-specific fluorochrome staining with 4'-6-diamidino-2-phenylindole (DAPI showed that the heterochromatin was rich in AT base pairs (DAPI+ except for pair 13, which was chromomycin A3 (CMA3 positive indicating an excess of GC base pairs. Our data also suggests that there was variation in heterochromatin base composition.

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

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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.

A subfertile patient diagnosed with testicular carcinoma in situ by immunocytological staining for AP-2gamma in semen samples

DEFF Research Database (Denmark)

Hoei-Hansen, C E; Rajpert-De Meyts, E; Carlsen, E

2005-01-01

patients, including patients with testicular cancer and subfertility. Cells positive for AP-2gamma were found only in semen samples from patients diagnosed a priori with testicular neoplasms and, surprisingly, in a 23 year old control subject with oligozoospermia and no symptoms of a germ cell tumour......, simple method based on immunocytological staining of a semen sample for AP-2gamma, a novel marker for CIS. The value of this method for diagnostic use in the clinic requires further careful validation in a large series of patients and controls, but the preliminary results are promising....

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...

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

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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.

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.

Evaluation of MODS Culture in the Diagnosis of Pulmonary Tuberculosis

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Z Aminzadeh

2012-05-01

Full Text Available

<strong>Background and Objectivesstrong>

Culture of M. tuberculosis is the golden standard for the diagnosis of TB which is a much more sensitive test than Smear examination. There is a strong need to use the new assays in order to speed up diagnostic methods. The aim of this research was to determine the evaluation of Microscopic Observation Drug Susceptibility culture in pulmonary tuberculosis in comparison with Ziehl-Neelsen stain and Lowenstein-Jensen culture of sputum.

<strong>Methods>

The research method was a Cross-sectional (diagnostic test and the technique was observational-interview type. If the patient's history revealed clinical criteria compatible with TB and the infectious specialist’s judgment was that of "TB suspected case, the patient was considered a pulmonary TB suspect. Then, in addition to sputum Ziehl-Neelsen stain and culture for Lowenstein-Jensen, we carried out MODS culture as well.

<strong>Results>

100 patients (48 male, 52 female with mean age of 52.9 ± 21.83 were evaluated. During sputum examination, 40% were Ziehl-Neelsen stain positive while 30% had positive sputum culture for Mycobacterium Tuberculosis in Lowenstein-Jensen and 47% had positive MODS culture. In comparison with sputum smear and Lowenstein-Jensen culture, MODS had a sensitivity of 82.5% and 86%, specificity of 77% and 70%, positive predictive value of 70% and 55%, negative predictive value of 86% and 92%, respectively.

<strong>Conclusion>

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

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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.

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

Chromogenic In Situ Hybridization and p16/Ki67 Dual Staining on Formalin-Fixed Paraffin-Embedded Cervical Specimens: Correlation with HPV-DNA Test, E6/E7 mRNA Test, and Potential Clinical Applications

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Roberta Zappacosta

2013-01-01

Full Text Available Although HPV-DNA test and E6/E7 mRNA analyses remain the current standard for the confirmation of human papillomavirus (HPV infections in cytological specimens, no universally adopted techniques exist for the detection of HPV in formalin-fixed paraffin-embedded samples. Particularly, in routine laboratories, molecular assays are still time-consuming and would require a high level of expertise. In this study, we investigated the possible use of a novel HPV tyramide-based chromogenic in situ hybridization (CISH technology to locate HPV on tissue specimens. Then, we evaluate the potential usefulness of p16INK4a/Ki-67 double stain on histological samples, to identify cervical cells expressing HPV E6/E7 oncogenes. In our series, CISH showed a clear signal in 95.2% of the specimens and reached a sensitivity of 86.5%. CISH positivity always matched with HPV-DNA positivity, while 100% of cases with punctated signal joined with cervical intraepithelial neoplasia grade 2 or worse (CIN2+. p16/Ki67 immunohistochemistry gave an interpretable result in 100% of the cases. The use of dual stain significantly increased the agreement between pathologists, which reached 100%. Concordance between dual stain and E6/E7 mRNA test was 89%. In our series, both CISH and p16INK4a/Ki67 dual stain demonstrated high grade of performances. In particular, CISH would help to distinguish episomal from integrated HPV, in order to allow conclusions regarding the prognosis of the lesion, while p16INK4a/Ki67 dual stain approach would confer a high level of standardization to the diagnostic procedure.

Chromogenic In Situ Hybridization and p16/Ki67 Dual Staining on Formalin-Fixed Paraffin-Embedded Cervical Specimens: Correlation with HPV-DNA Test, E6/E7 mRNA Test, and Potential Clinical Applications

Science.gov (United States)

Zappacosta, Roberta; Colasante, Antonella; Viola, Patrizia; D'Antuono, Tommaso; Lattanzio, Giuseppe; Capanna, Serena; Gatta, Daniela Maria Pia; Rosini, Sandra

2013-01-01

Although HPV-DNA test and E6/E7 mRNA analyses remain the current standard for the confirmation of human papillomavirus (HPV) infections in cytological specimens, no universally adopted techniques exist for the detection of HPV in formalin-fixed paraffin-embedded samples. Particularly, in routine laboratories, molecular assays are still time-consuming and would require a high level of expertise. In this study, we investigated the possible use of a novel HPV tyramide-based chromogenic in situ hybridization (CISH) technology to locate HPV on tissue specimens. Then, we evaluate the potential usefulness of p16INK4a/Ki-67 double stain on histological samples, to identify cervical cells expressing HPV E6/E7 oncogenes. In our series, CISH showed a clear signal in 95.2% of the specimens and reached a sensitivity of 86.5%. CISH positivity always matched with HPV-DNA positivity, while 100% of cases with punctated signal joined with cervical intraepithelial neoplasia grade 2 or worse (CIN2+). p16/Ki67 immunohistochemistry gave an interpretable result in 100% of the cases. The use of dual stain significantly increased the agreement between pathologists, which reached 100%. Concordance between dual stain and E6/E7 mRNA test was 89%. In our series, both CISH and p16INK4a/Ki67 dual stain demonstrated high grade of performances. In particular, CISH would help to distinguish episomal from integrated HPV, in order to allow conclusions regarding the prognosis of the lesion, while p16INK4a/Ki67 dual stain approach would confer a high level of standardization to the diagnostic procedure. PMID:24369532

Flat mount preparation for observation and analysis of zebrafish embryo specimens stained by whole mount in situ hybridization.

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Cheng, Christina N; Li, Yue; Marra, Amanda N; Verdun, Valerie; Wingert, Rebecca A

2014-07-17

The zebrafish embryo is now commonly used for basic and biomedical research to investigate the genetic control of developmental processes and to model congenital abnormalities. During the first day of life, the zebrafish embryo progresses through many developmental stages including fertilization, cleavage, gastrulation, segmentation, and the organogenesis of structures such as the kidney, heart, and central nervous system. The anatomy of a young zebrafish embryo presents several challenges for the visualization and analysis of the tissues involved in many of these events because the embryo develops in association with a round yolk mass. Thus, for accurate analysis and imaging of experimental phenotypes in fixed embryonic specimens between the tailbud and 20 somite stage (10 and 19 hours post fertilization (hpf), respectively), such as those stained using whole mount in situ hybridization (WISH), it is often desirable to remove the embryo from the yolk ball and to position it flat on a glass slide. However, performing a flat mount procedure can be tedious. Therefore, successful and efficient flat mount preparation is greatly facilitated through the visual demonstration of the dissection technique, and also helped by using reagents that assist in optimal tissue handling. Here, we provide our WISH protocol for one or two-color detection of gene expression in the zebrafish embryo, and demonstrate how the flat mounting procedure can be performed on this example of a stained fixed specimen. This flat mounting protocol is broadly applicable to the study of many embryonic structures that emerge during early zebrafish development, and can be implemented in conjunction with other staining methods performed on fixed embryo samples.

Automatic nuclei segmentation in H&E stained breast cancer histopathology images.

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Mitko Veta

Full Text Available The introduction of fast digital slide scanners that provide whole slide images has led to a revival of interest in image analysis applications in pathology. Segmentation of cells and nuclei is an important first step towards automatic analysis of digitized microscopy images. We therefore developed an automated nuclei segmentation method that works with hematoxylin and eosin (H&E stained breast cancer histopathology images, which represent regions of whole digital slides. The procedure can be divided into four main steps: 1 pre-processing with color unmixing and morphological operators, 2 marker-controlled watershed segmentation at multiple scales and with different markers, 3 post-processing for rejection of false regions and 4 merging of the results from multiple scales. The procedure was developed on a set of 21 breast cancer cases (subset A and tested on a separate validation set of 18 cases (subset B. The evaluation was done in terms of both detection accuracy (sensitivity and positive predictive value and segmentation accuracy (Dice coefficient. The mean estimated sensitivity for subset A was 0.875 (±0.092 and for subset B 0.853 (±0.077. The mean estimated positive predictive value was 0.904 (±0.075 and 0.886 (±0.069 for subsets A and B, respectively. For both subsets, the distribution of the Dice coefficients had a high peak around 0.9, with the vast majority of segmentations having values larger than 0.8.

Automatic nuclei segmentation in H&E stained breast cancer histopathology images.

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Veta, Mitko; van Diest, Paul J; Kornegoor, Robert; Huisman, André; Viergever, Max A; Pluim, Josien P W

2013-01-01

The introduction of fast digital slide scanners that provide whole slide images has led to a revival of interest in image analysis applications in pathology. Segmentation of cells and nuclei is an important first step towards automatic analysis of digitized microscopy images. We therefore developed an automated nuclei segmentation method that works with hematoxylin and eosin (H&E) stained breast cancer histopathology images, which represent regions of whole digital slides. The procedure can be divided into four main steps: 1) pre-processing with color unmixing and morphological operators, 2) marker-controlled watershed segmentation at multiple scales and with different markers, 3) post-processing for rejection of false regions and 4) merging of the results from multiple scales. The procedure was developed on a set of 21 breast cancer cases (subset A) and tested on a separate validation set of 18 cases (subset B). The evaluation was done in terms of both detection accuracy (sensitivity and positive predictive value) and segmentation accuracy (Dice coefficient). The mean estimated sensitivity for subset A was 0.875 (±0.092) and for subset B 0.853 (±0.077). The mean estimated positive predictive value was 0.904 (±0.075) and 0.886 (±0.069) for subsets A and B, respectively. For both subsets, the distribution of the Dice coefficients had a high peak around 0.9, with the vast majority of segmentations having values larger than 0.8.

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).

Light microscopy with differential staining techniques for the characterisation and discrimination of insects versus marine arthropods processed animal proteins.

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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.

Electromagnetic radiation from positive-energy bound electrons in the Coulomb field of a nucleus at rest in a strong uniform magnetic field

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Arsenyev, S. A.; Koryagin, S. A., E-mail: koryagin@appl.sci-nnov.ru [Russian Academy of Sciences, Institute of Applied Physics (Russian Federation)

2012-06-15

A classical analysis is presented of the electromagnetic radiation emitted by positive-energy electrons performing bound motion in the Coulomb field of a nucleus at rest in a strong uniform magnetic field. Bounded trajectories exist and span a wide range of velocity directions near the nucleus (compared to free trajectories with similar energies) when the electron Larmor radius is smaller than the distance at which the electron-nucleus Coulomb interaction energy is equal to the mechanical energy of an electron. The required conditions occur in magnetic white dwarf photospheres and have been achieved in experiments on production of antihydrogen. Under these conditions, the radiant power per unit volume emitted by positive-energy bound electrons is much higher than the analogous characteristic of bremsstrahlung (in particular, in thermal equilibrium) at frequencies that are below the electron cyclotron frequency but higher than the inverse transit time through the interaction region in a close collision in the absence of a magnetic field. The quantum energy discreteness of positive-energy bound states restricts the radiation from an ensemble of bound electrons (e.g., in thermal equilibrium) to nonoverlapping spectral lines, while continuum radiative transfer is dominated by linearly polarized bremsstrahlung.

Comparison of modified Chicago sky blue stain and potassium hydroxide mount for the diagnosis of dermatomycoses and onychomycoses.

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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.

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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.

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.

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.

Assessment of Proliferative Potential of Odontogenic Keratocyst and Dentigerous Cyst using Podoplanin: An Immunohistochemical Study.

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Gupta, Sandeep; Paliwal, Aparna; Choudaha, Nidhi; Gupta, Anish; Rao, Prashant; Grover, Shekhar

2017-12-01

Odontogenic cysts are commonly encountered lesions among head and neck pathologies. Odontogenic keratocyst (OKC) has unique features of recurrence and local aggressiveness. Podoplanin (PDP) is a lymphatic endothelial marker and is shown to be expressed in a variety of tissues. Hence, we planned to assess the significance of PDP in OKC and dentigerous cyst (DC). The present study included assessment of immunoexpression of PDP in OKC and DC. Twenty specimens each of OKC and DC were included in the present study and were stained with D2-40 antibody. All the sections were analyzed and were categorized as negative staining, weakly positive staining, and strongly positive staining. All the results were analyzed by Statistical Package for the Social Sciences (SPSS) software. We detected PDP-positive staining in the cell membrane and cytoplasm of the cells of basal cell layer and supra-basal cell layers. In DC cases, we observed positive staining only in cases associated with inflammation. Podoplanin does play a significant role in enhancing the local invasive and neoplastic properties of OKC. Podoplanin expression in OKC is potentially associated with moderate invasive nature of the neighboring structures.

Urgensi Redesain Kebijakan dan Sistem Penyelenggaraan Studi Keislaman pada Peskam STAIN Samarinda

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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

High-definition hematoxylin and eosin staining in a transition to digital pathology

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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.

Identification and characterization of microsporidia from fecal samples of HIV-positive patients from Lagos, Nigeria.

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Oladele Teslim Ojuromi

Full Text Available BACKGROUND: Microsporidia are obligate intracellular parasites that infect a broad range of vertebrates and invertebrates. They have been increasingly recognized as human pathogens in AIDS patients, mainly associated with a life-threatening chronic diarrhea and systemic disease. However, to date the global epidemiology of human microsporidiosis is poorly understood, and recent data suggest that the incidence of these pathogens is much higher than previously reported and may represent a neglected etiological agent of more common diseases indeed in immunocompetent individuals. To contribute to the knowledge of microsporidia molecular epidemiology in HIV-positive patients in Nigeria, the authors tested stool samples proceeding from patients with and without diarrhea. METHODOLOGY/PRINCIPAL FINDINGS: Stool samples from 193 HIV-positive patients with and without diarrhea (67 and 126 respectively from Lagos (Nigeria were investigated for the presence of microsporidia and Cryptosporidium using Weber's Chromotrope-based stain, Kinyoun stain, IFAT and PCR. The Weber stain showed 45 fecal samples (23.3% with characteristic microsporidia spores, and a significant association of microsporidia with diarrhea was observed (O.R. = 18.2; CI: 95%. A similar result was obtained using Kinyoun stain, showing 44 (31,8% positive samples with structures morphologically compatible with Cryptosporidium sp, 14 (31.8% of them with infection mixed with microsporidia. The characterization of microsporidia species by IFAT and PCR allowed identification of Enterocytozoon bieneusi, Encephalitozoon intestinalis and E. cuniculi in 5, 2 and 1 samples respectively. The partial sequencing of the ITS region of the rRNA genes showed that the three isolates of E.bieneusi studied are included in Group I, one of which bears the genotype B. CONCLUSIONS/SIGNIFICANCE: To our knowledge, this is the first report of microsporidia characterization in fecal samples from HIV-positive patients from

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.

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.)

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.

μCT of ex-vivo stained mouse hearts and embryos enables a precise match between 3D virtual histology, classical histology and immunochemistry

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