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PubMed1986

Identification of human milk kappa-casein on polyacrylamide gels by differential staining with Ethyl-Stains-all and chymosin sensitivity.

Ethyl-Stains-all (ESA), a cationic carbocyanine dye that stains phosphorylated, sialylated, and unmodified proteins differentially, was used to stain a human casein fraction enriched for its kappa-casein-like characteristics. The staining properties and chymosin sensitivity of this fraction were compared with those of human milk and bovine casein proteins. Phosphorylated human and bovine beta caseins stained blue with ESA. The sialic acid-containing bovine kappa-casein stained blue-green. The human kappa-like fraction was enriched for a protein that stained blue-green with ESA. Both bovine kappa-casein and the human blue-green-staining protein were susceptible to chymosin digestion at lower concentrations of chymosin than that required for digestion of beta-caseins. In each case, following chymosin digestion, a green-staining peptide of lower molecular weight replaced the original protein and para-kappa-casein was formed. Identification of human kappa-casein on SDS-polyacrylamide gels was based on its differential staining with ESA and chymosin sensitivity with respect to beta-casein.

Animals↗
PubMed1991

[Effect of heat-staining procedure on the gram staining properties of mycobacteria].

Since the establishment of Gram stain by H.C.Y. Gram in 1884, it has been widely and routinely used as an aid for differentiation of bacteria. The bacteria are divided into three categories by the staining properties; Gram-positive, -negative, and -indefinite. All the text books in the world describe that mycobacteria such as M. tuberculosis are Gram-positive. By the merest chance, however, it was found that M. lepraemurium grown in tissues was not stained by the routinely used Gram staining method. Therefore, we tried to stain some of the mycobacteria by the Gram staining procedure which is widely used at present. The results obtained indicated that the mycobacteria tested were divided into three groups; the unstainable group such as M. leprae and M. lepraemurium, the Gram-positive and difficult-to-stain group which involves such slow growing mycobacteria as M. tuberculosis, M. avium, and M. intracellulare, and the Gram-indefinite group which contains such rapid growing mycobacteria as M. phlei, M. smegmatis, and M. chelonae. However, if Gram stain is carried out by the heating procedure at the first staining step, all the mycobacteria would become Gram-positive. Therefore, we emphasize that Gram staining of mycobacteria should be performed by the heating procedure.

Hot Temperature↗
PubMed2005

[The comparison of HE staining and giemsa staining in detection of eosinophilic granulocytes in conjunctiva scroping].

PURPOSE: To find the better method of eosinophilic granulocytes staining by comparison between HE staining and Giemsa staining METHODS: Twenty conjunctivitis patients(40 eyes) were studied. The infected palpabral conjunctiva of both eyes were scroped and stained separately with Giemsa staining and HE staining. RESULT: Under microscope, eosinophilic granulocytes were more clear by Giemsa staining than HE staining. CONCLUSION: In showing eosinophilic granulocytes, Giemsa staining was better than HE staining.

Azure Stains↗
PubMed2000

Endotracheal intubation at birth for preventing morbidity and mortality in vigorous, meconium-stained infants born at term.

BACKGROUND: On the basis of evidence from non-randomised studies, it has been recommended that all babies born through thick meconium should have their tracheas intubated so that suctioning of their airways can be performed. The aim is to reduce the incidence and severity of meconium aspiration syndrome. However, for term babies who are vigorous at birth endotracheal intubation may be both difficult and unnecessary. OBJECTIVES: To determine if endotracheal intubation and suction of the airways at birth in vigorous term meconium-stained babies is more beneficial than routine resuscitation including aspiration of the oro-pharynx. SEARCH STRATEGY: The search was made from Oxford Database of Perinatal Trials, the Neonatal Trials Registry of the Cochrane Neonatal Collaborative Review Group and information obtained from knowledgeable practising neonatologists. SELECTION CRITERIA: Randomised trials which compared a policy of routine vs no (or selective) use of endotracheal intubation and aspiration in the immediate management of vigorous term meconium-stained babies at birth. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including mortality, meconium aspiration syndrome, other respiratory conditions, pneumothorax, need for oxygen supplementation, stridor, convulsions and hypoxic-ischaemic encephalopathy were abstracted and analysed using Revman 3.1.1. MAIN RESULTS: Four randomised controlled trials of endotracheal intubation at birth in vigorous term meconium-stained babies were identified. Meta-analysis of these trials does not support routine use of endotracheal intubation at birth in vigorous meconium-stained babies to reduce mortality, meconium aspiration syndrome, other respiratory symptoms or disorders, pneumothorax, oxygen need, stridor, HIE and convulsions. REVIEWER'S CONCLUSIONS: Routine endotracheal intubation at birth in vigorous term meconium-stained babies has not been shown to be superior to routine resuscitation including oro-pharyngeal suction. This procedure cannot be recommended for vigorous infants until more research is available.

Humans↗
PubMed1995

Dietary staining in vitro by mouthrinses as a comparative measure of antiseptic activity and predictor of staining in vivo.

Extrinsic staining of teeth is a side-effect of some antiseptic mouthrinses. However, few of the many rinse products available to the general public have been investigated for their propensity to cause staining. Dietary factors play an aetiological role in staining and have been used in vitro to study and compare the activity of rinses. The aim of this study was to assess rinse products for staining in vitro and, through the staining reaction, to compare the activity of products containing the same ingredients. Perspex blocks, with or without saliva pretreatment, were soaked in rinses for 2 min, washed and placed in a standard tea solution for 60 min and then the optical density (OD) read on a spectrophotometer. The cycle was repeated 10 times for saliva and 17 times for no saliva specimens or until the maximum OD was exceeded. A series of three separate experiments was performed by this method. The maximum OD was not exceeded by any product before seven passages and therefore data were compared at six passages. For most products OD increased with saliva pretreatment. Some cetylpyridinium chloride (CPC) rinses stained comparably to a chlorhexidine rinse. CPC rinses, most of which contained the same concentration of the antiseptic, varied considerably in their propensity to induce staining and one was little different to water controls. A 0.1% chlorhexidine rinse stained slightly more than a 0.2%. A phenolic/essential oil product produced some staining but zinc, triclosan and other essential oil rinses did not stain.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗
PubMed1989

Standardized thionin-eosin stain in bronchial cytology. A substitute for hematoxylin-eosin Y staining.

A standardized thionin-eosinic acid stain was developed as a quick and highly reproducible staining method for bronchial cytology. Bronchial smears and paraffin-embedded sputum samples were stained with thionin-eosin and with the conventional hematoxylin-eosin Y. Spectral absorption characteristics and staining intensity of thionin-eosin-stained cells were investigated by means of cytophotometry. The staining pattern of thionin-eosin is very close to that of the hematoxylin-eosin stain; the contrast between nucleus and cytoplasm is significantly higher for thionin-eosin. Thionin-eosin can be used for "dye-fixation" of cytologic smears and tissue imprints. Blueing and differentiation (as for hematoxylin-eosin) is not required for thionin-eosin; thus, fixation and staining can be performed within two minutes. The spectral absorption characteristics of thionin-eosin allow reliable automated cytophotometric discrimination of cell nuclei and cytoplasm. The standardized thionin-eosin stain is recommended as a substitute for the hematoxylin and eosin stain in bronchial cytology.

Bronchi↗
PubMed1996

[Detection of bacterial vaginosis in gram stained vaginal smears and papanicolaou stained cervical smears].

OBJECTIVE: To determine the sensitivity and specificity of diagnosis of bacterial vaginosis (BV) by papanicolaou (PAP)-stained cervical smears and gram-stained vaginal smears according to clinical criteria. METHODS: In the prospective study of 196 non-pregnant women of child-bearing age, 94 were clinically diagnosed to have BV. Gram-stained vaginal smears and PAP-stained cervical smears were collected from all enrolled patients. RESULTS: In comparison with clinical diagnosis of BV, the sensitivity and specificity of gram-stained vaginal smears and PAP-stained cervical smears were 94.7% (89/94), 98.0% (100/102) 85.1% (80/94), and 95.1% (97/102). The characteristics of BV in gram-stained vaginal smears and PAP-stained cervical smears included small bacteria of Gardnerella and bacteroides morphotypes and curved gram variable rods and absence of bacteria of Lactobacillus morphotypes. CONCLUSION: It is possible to screen and manage BV according to Gram-stained vaginal smears or PAP-stained cervical smears.

Adult↗
PubMed1998

Effect of prolonged staining with hematoxylin on detecting Helicobacter pylori in hematoxylin-eosin-stained gastric mucosa.

Helicobacter pylori (H. pylori) is identifiable in hematoxylin-eosin (HE)-stained preparations, particularly under a prolonged hematoxylin staining condition. The staining intensity of H. pylori bodies was increased when the hematoxylin staining period was doubled from 5 min, the routine period of Meyer's hematoxylin staining, to 10 min. The staining intensity of the background epithelial nuclei was only mildly increased. Hematoxylin-eosin staining employing a prolonged hematoxylin staining period gave detectability of H. pylori comparable to May-Giemsa and immunoperoxidase staining. The modified HE method is thus very useful and practical for identifying H. pylori in routine gastric biopsy specimens.

Adult↗
PubMed1988

One-bath trichrome staining: investigation of a general mechanism based on a structure-staining correlation analysis.

Dye pairs of contrasting colours were selected from acid dyes of varied chemical characteristics. The 44 dye pairs were investigated in a one-bath trichrome staining system in which the dye-baths were strongly acid. Dye concentrations, concentration ratios and staining times were varied for each dye pair. Thirty dye pairs stained collagen fibres distinctly different colours to muscle cytoplasm, while 14 dye pairs gave muddy, non-selective staining. Comparison of dye structures showed that in selective pairs the larger dye always stained the collagen fibres, with cytoplasm being coloured by the smaller species. With 28/30 of the selective dye pairs the differences in anionic weights of the dyes was greater than 200. However, in dye pairs giving non-selective staining, the anionic weights of the members of 13/14 of the dye pairs differed by less than 200. As no other structural feature correlated so clearly with selectivity, it was concluded that the selectivity of one-bath trichromes is diffusion-rate controlled, involving the interaction of differentially permeable tissue sites (collagen being more permeable than muscle cytoplasm) with dyes diffusing at different rates (large dyes slower than small). In keeping with this, lengthening staining times reduced staining selectivity. The rate control mechanism suggested a rational trouble-shooting guide for one-bath trichromes, encompassing such practical factors as dye concentration, embedding medium, fixative, dye-bath pH, section thickness and staining time.

Animals↗
PubMed2006

ProteomIQ blue, a potent post-stain for the visualization and subsequent mass spectrometry based identification of fluorescent stained proteins on 2D-gels.

Manual spot excision for protein identification from fluorescent stained two-dimensional (2-D) gels is hard to accomplish. Here, we explore the use of ProteomIQ Blue as a post-stain method for the visualization of fluorescent stained/labeled proteins. We show that ProteomIQ Blue post-staining is almost as sensitive as staining with SYPRO Ruby or cyanine dyes alone. More than 90% of the protein spots that are stained with the fluorescent stains are still detectable with ProteomIQ Blue. In protein identification by mass spectrometry, ProteomIQ Blue post-stained spots provide high sensitivity and high protein sequence coverage of the peptide mass maps in both MALDI-TOF-MS and ESI-MS/MS analyses. In conclusion, post-staining of fluorescent stained gels with ProteomIQ Blue provides a facile and a powerful method to achieve quantitative protein analysis as well as protein identification in the same semianalytical gel without requiring sophisticated/expensive robotic equipment.

Bacterial Proteins↗
PubMed2003

Traditional staining for routine diagnostic pathology including the role of tannic acid. 1. Value and limitations of the hematoxylin-eosin stain.

The components of the hematoxylin and eosin (H & E) stain (i.e. hemalum and eosin Y), their contributions to the typical staining pattern, and the reasons why the H & E stains are the preferred oversight stains for routine diagnostic histopathology are discussed. The essential diagnostic significance of effective nuclear staining by hemalum, providing information on nuclear morphology and texture, is emphasized; as is the ironic advantage for routine diagnostic histopathology of the limited range of colors provided by H & E staining, that allows recognition of significant features under low microscopic magnifications. Standardization of hemalum is considered, along with probable reasons why users show resistance to such a concept. Counterstaining with anionic (acid) dyes is discussed, as is the important phenomenon of contrast. The particular advantages and disadvantages of eosin Y and phloxin B as counterstains to hemalum are outlined. The concept of an "ideal routine histological stain" is considered, and H & E is compared to such an ideal case. Finally, deficiencies of H & E staining are discussed, and a program to develop an improved oversight stain is introduced.

Animals↗
PubMed1995

Immunohistochemical staining of ras p21: staining in benign and malignant prostate tissue.

A total of 124 specimens of prostate tissue (25 normal prostate, 41 benign prostatic hyperplasia, 58 adenocarcinoma) was immunostained for ras p21 using a commercially available monoclonal antibody directed against a peptide sequence conserved among all members of the ras gene family. Of normal prostate specimens, 76% showed no staining while the remainder showed only weak epithelial (glandular) staining. No significant stromal staining was noted in any normal prostate specimen. In contrast most benign prostatic hyperplasia specimens showed abundant staining. Epithelial staining was observed in 88% and stromal staining in 73% of specimens. A majority of prostate carcinoma specimens also stained, with 62% and 36% showing epithelial and stromal staining, respectively. No association was noted between staining and either tumor grade or clinical stage. These data argue against any clinical usefulness of immunostaining for ras p21 in the diagnosis or grading of prostate cancer.

Adenocarcinoma↗
PubMed1983

Quantitative immunohistochemistry: a comparison of microdensitometric analysis of unlabeled antibody peroxidase-antiperoxidase staining and of microfluorometric analysis of indirect fluorescent antibody staining for nicotinamide adenosine dinucleotide phosphate (NADPH)-cytochrome c (P-450) reductase in rat liver.

The intralobular distribution of nicotinamide adenine dinucleotide phosphate (NADPH)-cytochrome c (P-450) reductase (NADPH:ferricytochrome oxidoreductase, EC 1.6.2.4) in rat liver has been investigated by means of two quantitative immunohistochemical techniques: microdensitometric quantitation of unlabeled antibody peroxidase-antiperoxidase staining and microfluorometric analysis of indirect fluorescent antibody staining. Utilizing sheep antiserum elicited against NADPH-cytochrome c (P-450) reductase that had been isolated and purified to apparent homogeneity from rat liver microsomes, the reductase was detected within hepatocytes throughout the liver. However, differences in the intensity of staining of hepatocytes within different regions of the liver lobule were readily apparent after completion of both immunohistochemical staining procedures. These visual findings were verified by microdensitometric and microfluorometric analyses of immunohistochemical staining, both of which revealed that approximately the same degree of staining for NADPH-cytochrome c (P-450) reductase was produced within the centrilobular and midzonal regions of the liver lobule, whereas periportal hepatocytes were stained with significantly less intensity. These results demonstrate that the application of either microdensitometry in conjunction with unlabeled antibody peroxidase-antiperoxidase staining or microfluorometry after indirect fluorescent antibody staining can be used to quantitatively determine the intratissue distributions of antigens.

Animals↗
PubMed1982

Basic dyes for the staining of DNA in mammalian tissues and absorption spectra of stained nuclei in the visible light.

This communication presents informations on the staining of DNA with basic dyes, such as ethyl violet, Janus green, a new red dye obtained from Janus green, and Giemsa, belonging to different chemical groups. It has been found that DNA of tissue sections from which RNA has been extracted selectively with cold concentrated phosphoric acid can be stained with aqueous solutions of these dyes. Further, DNA of tissue sections from which RNA has been extracted and the sections then hydrolysed in 6 N HCl at 28 degrees C for 15 min can also be stained with aqueous solutions of ethyl violet, Janus green, and Giemsa. Moreover, tissue sections that have been hydrolysed in 6 N HCl and then stained with aqueous solutions of these dyes, including the new red dye obtained from Janus green, reveal well-stained nuclei. The absorption spectra of nuclei stained with aqueous solutions of ethyl violet, Janus green and Giemsa, following the above stated three procedures of staining have been presented. Absorption data of nuclei stained with the new red dye after Feulgen hydrolysis of tissue sections as well as those of nuclei in tissue sections from which RNA has been extracted have also been presented herein. The implications of these findings have been discussed.

Animals↗
PubMed1990

Silver staining of interphase nucleolar organizer regions in cytologic smears previously stained by the Papanicolaou and May-Grünwald-Giemsa techniques.

Malignant cells are known to display a distinctive argyrophilic nucleolar organizer region (Ag-NOR) protein distribution compared with distribution in their benign counterparts. In this study, Ag-NOR staining was applied to smears from reactive and malignant serous effusions. The smears were either unstained or had been previously stained with the Papanicolaou or May-Grünwald-Giemsa (MGG) technique. The purpose of this study was to assess the feasibility and usefulness of Ag-NOR staining in diagnostic cytopathology and to set up a procedure that could be used on prestained smears in retrospective studies. The one-step silver staining method was applied to prestained smears after a weak destaining step in trichloracetic acid and to unstained smears after a postfixation step in Carnoy's fluid. Positive results were observed on both destained and unstained preparations, MGG-stained smears showing a better visualization of the silver deposits than Papanicolaou-stained smears. Major differences in size, shape, and distribution patterns of the Ag-NOR-positive granules were observed between neoplastic and reactive cells. Furthermore, the mean number of silver-stained dots per cell was significantly higher in malignant cells (33.32 +/- 2.23) than in reactive mesothelial cells (9.71 +/- 0.66). These data indicate that this Ag-NOR technique can be profitably applied to prestained cytologic smears to assist in the diagnosis of malignancy and that the technique has the advantage that cellular morphology and silver staining can be evaluated on the same slide.

Ascitic Fluid↗
PubMed2006

Nicotinamide adenine dinucleotide staining immediately following radio frequency ablation of renal tumors-is a positive stain synonymous with ablative failure?

PURPOSE: Nicotinamide adenine dinucleotide diaphorase staining is arguably the standard for assessing tissue viability following radio frequency ablation, yet the accuracy of this test is questionable. Thus, it is imperative to examine the reliability of nicotinamide adenine dinucleotide to predict ablation success. To evaluate the observation that immediate nicotinamide adenine dinucleotide staining may not be clinically reliable, we compared results from immediate post-ablation biopsies of renal tumors to radiological and clinical followup. MATERIALS AND METHODS: Laparoscopic radio frequency ablation was performed in 9 patients and 10 tumors using a temperature modulated radio frequency system. Cold cup biopsies were taken immediately following ablation, and processed for nicotinamide adenine dinucleotide and hematoxylin and eosin staining. Patients were then followed using contrast enhanced computerized tomography at regular intervals. RESULTS: Median tumor size was 2.3 cm. Hematoxylin and eosin stain diagnosed 8 renal cell carcinomas and 2 angiomyolipomas. A quarter of the renal cell carcinomas and both angiomyolipomas stained positive for scattered nicotinamide adenine dinucleotide diaphorase activity immediately after RFA. Mean followup for the nicotinamide adenine dinucleotide positive tumors was 28.5 months (range 24 to 30) and for the nicotinamide adenine dinucleotide negative tumors was 25 months (range 18 to 30). There was no evidence of local tumor recurrence in any patient. CONCLUSIONS: Four lesions had nicotinamide adenine dinucleotide diaphorase activity on post-ablation biopsy suggesting retained viable tissue, yet there has been no recurrence during an average 2-year followup. While negative nicotinamide adenine dinucleotide staining is consistent with nonviability, these results suggest that false-positive staining can occur immediately following RFA, making the predictive value of positive nicotinamide adenine dinucleotide diaphorase staining unclear.

Adult↗