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PubMed1985

Comparison of the automicrobic system, acridine orange-stained smears, and gram-stained smears in detecting bacteriuria.

We compared the accuracy of the Gram-stained smear, the acridine orange-stained smear, and the AutoMicrobic system (AMS; Vitek Systems, Inc., Hazelwood, Mo.) in screening for bacteriuria, as detected by conventional cultures. For 1,024 clinical specimens, results with the acridine orange-stained smear and the Gram-stained smear were very similar. When read for the presence of one or more microorganisms or leukocytes per 20 oil immersion fields, both smears were highly sensitive (92.1 and 93.3%, respectively) and moderately specific (70.0 and 61.7%, respectively). Sensitivity was greater for specimens yielding greater than or equal to 10(5) CFU/ml (96.1 and 98.9%, respectively) than for those with 10(3) to 10(4) CFU/ml (81.4 and 78.0%, respectively). Preliminary classification based upon the tinctorial and morphological characteristics of the Gram-stained smear was compatible with culture results in nearly all cases. The accuracy of the Gram-stained smears was not influenced by special cleaning of the microscopic slides, or the level of expertise of the microscopist. For 715 specimens, the sensitivity of the AMS in detecting bacteriuria (91.5%) was very similar to that of the stained smears (92.1 and 95.7%, respectively), but the specificity was significantly higher (83.2% versus 42.6 and 70.0%). Detection of microorganisms by the AMS took an average of 6.3 +/- 3.0 h. These data suggest that the Gram-stained smear is easily interpreted, very sensitive, acceptably specific, and still the optimal rapid method for screening for bacteriuria in most clinical microbiology laboratories.

Acridine Orange↗

PubMed1986

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

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.

Brain↗

PubMed1983

Comparison of Papanicolaou staining and Feulgen staining for automated prescreening.

Feulgen staining is considered to be a quantitative DNA-specific cytochemical procedure. The applicability of this staining in high-resolution cytometry was tested in comparison with a regressive Papanicolaou staining. Papanicolaou-stained or Feulgen-stained intermediate and carcinoma cells selected by a cytologist were examined with a Zeiss scanning microscope photometer at 546 and 560 nm, respectively. After cell image segmentation and feature extraction, a statistical data evaluation was carried out by computer. Cell distributions with respect to four selected nuclear features demonstrated the influence of the staining procedure on cell feature measurements. The discriminatory power of the classification system as related to both staining procedures was studied using discriminant analysis. Using only nuclear features, a 7.3% improvement of the overall correct classification rate (from 85.0% to 92.3%) was achieved using Feulgen staining. The misclassification rate was simultaneously reduced by 50%. Using cytoplasmic as well as nuclear features, a 98% rate of correct classification was achieved.

Cell Nucleus↗

PubMedDate not supplied

Immunocytochemical staining of unstained versus previously stained cytologic preparations.

Immunoperoxidase (IP) staining of fine needle aspiration specimens may provide useful adjunct information in cytodiagnosis. IP may be used on unstained direct smears or cell block material if available. In the absence of such material, IP is sometimes used on previously stained (Papanicolaou) smears. Few studies have examined the adequacy of such preparations, however. We compared IP staining results on unstained, alcohol-fixed or air-dried smears versus alcohol-fixed, previously Papanicolaou stained smears and versus alcohol-fixed, decolorized, Papanicolaou-stained smears, using antibodies to keratin (CAM 5.2, AE1/AE3 and K903), epithelial membrane antigen, monoclonal carcinoembryonic antigen, desmin, muscle-specific actin (HHF-35), vimentin, leukocyte common antigen, B and T cell markers L26 and UCHL-1, and prostate-specific antigen. Seventy-five sets of smears were made from various neoplastic or nonneoplastic tissues received as surgical specimens, and antibodies were selected for anticipated positive staining. The results were graded in a semiquantitative fashion. All the methods gave comparable results regarding expected positive staining, background staining and preservation of cellularity. We conclude that these methods of smear preparation are of equal utility in adjunctive immunocytochemical studies.

Biopsy, Needle↗

PubMed1979

Copper stains and the syndrome of primary biliary cirrhosis. Evaluation of staining methods and their usefulness for diagnosis and trials of penicillamine treatment.

Use of Shikata's stain was found to be a sensitive method for the demonstration of copper-protein complexes, but use of the rhodamine stain may be more reliable. Interobserver agreement was slightly better with Shikata's stain. Hepatic copper increased with the histologic progression of chronic nonsuppurative destructive cholangitis (CNDC). All specimens that contained Mallory bodies and most specimens (91%) that contained bile yielded positive copper stains. No correlation was found between positive copper stains and other histologic features. A strongly positive copper stain aided in the diagnosis of CNDC. All specimens with negative copper stains contained less than 250 microgram of copper per gram of dry weight. From a strongly positive copper stain, the chemical copper content could not be predicted with certainty. Shikata's method seemed adequate to evaluate penicillamine treatment trials.

Adult↗

PubMed2000

Amnioinfusion for meconium-stained liquor in labour.

BACKGROUND: 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. OBJECTIVES: The objective of this review was to assess the effects of amnioinfusion for meconium-stained liquor on perinatal outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched. SELECTION CRITERIA: Randomised trials comparing amnioinfusion with no amnioinfusion for women in labour with moderate or thick meconium-staining of the amniotic fluid. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Ten 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 0.47, 95% confidence interval 0.24 to 0. 90); and a trend to reduced caesarean section overall (relative risk 0.83, 95% confidence interval 0.69 to 1.00). 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). REVIEWER'S CONCLUSIONS: Amnioinfusion is associated with improvements in perinatal outcome, particularly in settings where facilities for perinatal surveillance are limited. The trials reviewed are too small to address the possibility of rare but serious maternal adverse effects of amnioinfusion.

Amnion↗

PubMedDate not supplied

Pleural and pulmonary staining at inferior phrenic arteriography mimicking a tumor staining of hepatocellular carcinoma.

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.

Carcinoma, Hepatocellular↗

PubMed2004

Clinical evaluation of the stain removing ability of a whitening dentifrice and stain controlling system.

OBJECTIVES: This study aimed to evaluate the stain removing/controlling properties of a whitening dentifrice and a stain controlling system, in a parallel group, multiple use study. METHODS: Subjects (63), were stratified according to baseline stain scores and randomly allocated to a group, A (standard dentifrice), B (whitening/stain removing dentifrice) or C (whitening/stain removing dentifrice, tooth polish, tooth polisher and whitening/stain removing mouthwash). At each attendance, extrinsic stain was scored blind by a trained and calibrated examiner using the Shaw and Murray Stain Index, under constant lighting conditions. In addition, whiteness was measured using a colorimeter. RESULTS: The toothpaste regimens were well balanced at baseline (p=0.811). At 4 weeks, Regimens B and C had significantly less stain than A (p<0.05). Lower stain values were maintained/reduced for regimens B and C by week 6, with changes from baseline significantly greater than for Regimen A. When lightness was assessed by colorimeter, Regimen C was also found to be significantly whiter than Regimen A (p<0.05). CONCLUSIONS: It was concluded that both whitening regimens had a significant effect on reducing tooth staining and improving whiteness over the 6 week time period.

Adult↗

PubMed2005

Comparison of quantitative light-induced fluorescence (QLF) and digital imaging applied for the detection and quantification of staining and stain removal on teeth.

OBJECTIVES: This study compares the use of QLF with digital imaging in the detection and quantification of the development and removal of stain on teeth. METHODS: Two experimental phases, tooth staining and tooth whitening, conducted in vitro on labial 12 mm(2) enamel windows made on ten extracted bovine teeth, developed stains in 6-min cycles (2 min in each solution) using artificial saliva, chlorhexidine and tea solutions and removed them using sodium perborate monohydrate in 2-min cycle monitored at the end of each cycle with QLF (Inspektor Research Systems, NL) and digital photography (Fuji, Japan). The stain values were quantified as DeltaQ derived from QLF and DeltaE from digital imaging. This was observed by the two methods correlated with Pearson correlation coefficient (r). Regression equations (R(2)) were also obtained. RESULTS: For both staining and stain removal there was a statistically significant (p<0.01) reverse correlation between DeltaQ values for QLF (r=-0.924, R(2)=85.4%) and DeltaE values for digital imaging (r=-0.994, R(2)=98.8%), respectively. CONCLUSION: QLF showed a high correlation with digital imaging as a technique for detecting and monitoring tooth stains and tooth whitening in vitro. The potential for QLF with further development as a tool for monitoring staining and whitening of teeth may be possible in vivo in addition to the diagnostic ability for caries detection.

Animals↗

PubMed1995

Detection of bacterial vaginosis in wet mount, Papanicolaou stained vaginal smears and in gram stained smears.

In a prospective study of 107 women, bacterial vaginosis was clinically diagnosed in 34 women. Compared with clinical diagnosis of bacterial vaginosis, detection of clue cells in Papanicolaou stained vaginal smears showed a sensitivity of 88.2%, a specificity of 98.6%, a positive predictive value of 96.8% and a negative predictive value of 94.7%. The corresponding values for detection of bacterial vaginosis in Gram stained smears compared with the clinical diagnosis were 100%, 97.3%, 94.4% and 100%, respectively. Compared with clue cells in wet smears, identification of clue cells in Papanicolaou stained vaginal smears showed a Kappa index of 0.87 and compared with Gram stain criteria a Kappa index of 0.94. The correlation between Gram stain and Papanicolaou stained vaginal smears showed a Kappa index of 0.89. In contrast to the results of earlier investigators our studies indicate that the demonstration of clue cells in Papanicolaou stained vaginal smears correlate reasonably well with the conventional clinical criteria. However, the Gram stain method may be more reliable than the Papanicolaou method.

Female↗

PubMed1997

[Transferrin subtyping of human semen, semen stain, vaginal fluid and mixed stain using isoelectric focusing and immunoblotting].

Transferrin (Tf) is an important genetic marker for personal identification in forensic science. We examined the subtypes in human semen, semen stain, vaginal fluid and mixed stain using isoelectric focusing and immunoblotting method(IEFIB). The results showed that, Tf subtypes of 8 semen and semen stain samples stored at room temperature for up to 32 weeks were the same as those in sera. Tf subtypes of 17 vaginal fluid and 4 mixed stain containing few semen were not detected. Tf Subtyping of 1 mixed stain containing enough semen was successful by using IEFIB. These results indicate that Tf is stable in semen stain. The Tf subtyping in semen stain and mixed stain containing enough semen can be carried out by using IEFIB method.

Exudates and Transudates↗

PubMed1992

Morphometry of cupromeronic blue-stained proteoglycan molecules in animal corneas, versus that of purified proteoglycans stained in vitro, implies that tertiary structures contribute to corneal ultrastructure.

Isolated, purified small chondroitin (dermatan) sulphate proteoglycans from corneas of cow and rabbit and cow sclera were stained with Cupromeronic blue in 'model' experiments. The lengths and thicknesses of the images were compared with those of the same proteoglycans stained in the tissue, using the critical electrolyte concentration principle to give specificity for sulphated proteoglycans, and keratanase 1 or chondroitinase ABC digestion to distinguish between chondroitin and keratan sulphate. Corrections for orientation of the stained glycan filaments within the section plane were made to convert the observed lengths to true average lengths. Observed lengths of stained chondroitin (dermatan) sulphate were greater than those of keratan sulphate, both in models and tissues, in agreement with published data from biochemical and rotary-shadowing studies, in both species. Corrected (true) average lengths of stained isolated chondroitin (dermatan) sulphate proteoglycans were slightly, but not significantly, longer than expected from rotary shadowing or biochemical measurements. Keratan sulphate lengths were similarly somewhat longer. The data support the idea that Cupromeronic blue acts as a scaffold that helps maintain polyanion shape against distortion on staining. Stained filaments in tissues were sometimes over twice the length of isolated stained proteoglycans, suggesting that 2 glycan chains were aligned end-to-end. Thicknesses of proteoglycan filaments suggested that at least 2 glycan chains were aligned side-by-side, both in models and in tissues. A scheme for proteoglycan tertiary structure in cornea is proposed, in which glycan chains may bridge collagen fibrils in duplexed forms similar to those observed in rotary shadowed preparations.

Animals↗

PubMed2000

A comparison of silver stain and SYPRO Ruby Protein Gel Stain with respect to protein detection in two-dimensional gels and identification by peptide mass profiling.

Proteomic projects are often focused on the discovery of differentially expressed proteins between control and experimental samples. Most laboratories choose the approach of running two-dimensional (2-D) gels, analyzing them and identifying the differentially expressed proteins by in-gel digestion and mass spectrometry. To date, the available stains for visualizing proteins on 2-D gels have been less than ideal for these projects because of poor detection sensitivity (Coomassie blue stain) or poor peptide recovery from in-gel digests and mass spectrometry (silver stain), unless extra destaining and washing steps are included in the protocol. In addition, the limited dynamic range of these stains has made it difficult to rigorously and reliably determine subtle differences in protein quantities. SYPRO Ruby Protein Gel Stain is a novel, ruthenium-based fluorescent dye for the detection of proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels that has properties making it well suited to high-throughput proteomics projects. The advantages of SYPRO Ruby Protein Gel Stain relative to silver stain demonstrated in this study include a broad linear dynamic range and enhanced recovery of peptides from in-gel digests for matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry.

Acrylic Resins↗

PubMedDate not supplied

Cytology of polychrome-stained equine synovial fluid smears. Comparison with clinical findings, histologic specimens, Wright-Giemsa-stained smears and outcome.

Polychrome-stained equine synovial fluid specimens from 34 normal joints and 129 joints with clinical abnormalities were examined cytologically. The smears from joints with abnormalities were categorized as within normal limits (4.7%), slight abnormality (27.9%), proliferative synovitis (21.7%), neutrophilic pattern (20.2%), elongated cell pattern (10.1%), other moderate to marked abnormality (11.6%) and unsatisfactory (3.9%). Cytologic abnormalities that were not restricted to a single category included spindle cells, crystals, stellate cells and cartilage fragments. Multinucleate cells and mononucleate cells with dense cytoplasm and a delicate periphery were seen in smears from cases with clinical diagnoses of osteochondrosis or fracture; interpretation of these cells as osteoclasts and their mononucleate precursors was supported by positive staining with tartrate-resistant acid phosphatase. Smears within the same cytologic category were not found to correspond with a single clinical diagnosis. The identification of several cytologic patterns in cases with the same clinical diagnosis suggests that multiple stages of disease were sampled. Except in cases with the cytologic neutrophilic pattern, there was not a consistent relationship between the histologic features in synovial biopsy specimens and the cytologic findings; the morphologic variation within synovial membrane sections and between sections from different locations was sometimes marked. When compared with air-dried, Wright-Giemsa-stained smears, the polychrome-stained smears were more sensitive in the detection of cytologic abnormalities and were less often falsely negative or unsatisfactory. Following surgery, cases with clinical diagnoses of osteochondrosis (29 cases) and fracture (25 cases) were analyzed according to clinical outcome and cytologic category. While 80% of the horses with proliferative synovitis in cytologic specimens were sound, only 67% of those with the elongated cell pattern, 50% of those with slight abnormality and 33% of those with other moderate to marked abnormality were sound. A statistically significant relationship (P less than .02) was found in cases with a diagnosis of osteochondrosis: animals with a proliferative synovitis pattern were almost three times as likely to be sound as compared to those with slight abnormality. These findings indicate that polychrome-stained equine synovial fluid smears (1) provide information that is different from that found in corresponding histologic sections and (2) are superior to air-dried, Wright-Giemsa-stained smears for cytologic examination. The polychrome-stained equine synovial fluid smears were found to provide information supportive of clinical, radiographic and prognostic data.

Animals↗

PubMed2000

Development of a mechanism-based, DNA staining protocol using SYTOX orange nucleic acid stain and DNA fragment sizing flow cytometry.

Accurate measurement of single DNA fragments by DNA fragment sizing flow cytometry (FSFC) depends upon precise, stoichiometric DNA staining by the intercalating dye molecules. In this study, we determined the binding characteristics of a commercially available 532 nm wavelength-excitable dye and used this information to develop a universal DNA staining protocol for DNA FSFC using a compact frequency-doubled Nd:YAG laser excitation source. Among twelve 532 nm wavelength-excitable nucleic acid staining dyes tested, SYTOX Orange stain showed the highest fluorescence intensity along with a large fluorescence enhancement upon binding to double-stranded DNA ( approximately 450-fold). Furthermore, using SYTOX Orange stain, accurate fragment-size-distribution histograms were consistently obtained without regard to the staining dye to base pair (dye/bp) ratio. A model describing two binding modes, intercalation (primary, yielding fluorescence) and external binding (secondary, involving fluorescence quenching), was proposed to interpret the performance of the dyes under different dye/bp ratios. The secondary equilibrium dissociation constant was found to be the most critical parameter in determining the sensitivity of each fluorophore to the staining dye/bp ratio. The measurements of both equilibrium dissociation constants provided us with a theoretical framework for developing a universal protocol which was successfully demonstrated over a wide range of DNA concentrations on a compact flow cytometer equipped with a frequency-doubled, diode-pumped, solid-state Nd:YAG laser for rapid and sensitive DNA fragment sizing.

Bacteriophage lambda↗

PubMedDate not supplied

Application of stains-all staining to the analysis of axonemal tubulins: identification of beta-tubulin and beta-isotubulins.

The cationic dye, Stains-all, is known to stain brain beta-tubulin blue and alpha-tubulin red (Serrano, L. et al. (1986) J. Biochem. Biophys. Methods 12, 281-287; Serrano, L. et al. (1989) Biochem. Int., 19, 235-246). The present experiments show that this stain can also be applied to detect beta-tubulin in axonemal tubulins from various sources such as cilia of protozoa, sperm flagella of echinoderm, and sperm flagella of mollusc. Furthermore, these experiments showed that it selectively stains isoforms of axonemal beta-tubulin blue following isoelectric focusing, whereas those of alpha-tubulin are stained red. These results indicate that Stains-all staining is a useful tool for electrophoretic analysis of axonemal tubulins.

Animals↗

PubMed1990

Staining of light-cured aesthetic resin restorative materials by different staining media: an in vitro study.

Aesthetic restorative resins have been characterised by clinical discolouration. New light-cured anterior and posterior resins are currently available to the profession and this study investigated the in vitro colour stability at 24 hours and 7 days of ten such resins in three different staining media. Disc specimens were prepared and stained at 37 degrees C and 100 per cent R. H. and then evaluated by double blind visual assessment. All the materials undergo some staining in coffee and red wine. Coca-Cola, however, does not cause any staining of the restorative resins. Adaptic II is the most stain-resistant material and most of the staining occurs within the first 24 hours after immersion in the staining media.

Bisphenol A-Glycidyl Methacrylate↗

PubMed1989

Automated Feulgen staining with a temperature-controlled staining machine.

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

Coloring Agents↗