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At least 685 records · Page 38Linked to original sources
PubMed1985

The median forehead flap revisited: the blood supply.

In 6 fresh cadavers, an injection study of the facial vessels with disulfine blue dye and Microfil demonstrated visualization of large-caliber vessels of the median forehead skin even when the supraorbital and supratrochlear vessels were interrupted. The results of the study would suggest that the median forehead flap can be elevated without incorporating the supratrochlear vessels, but the flap design should be reserved for those clinical situations where the pedicle must be extensively mobilized, e.g., reconstruction of the nasal tip and columella and the presence of a low-lying frontal hairline.

Angiography↗
PubMed1975

A methanol resorcin-fuchsin stain for elastic tissues and nuclei.

The staining properties of conventional ethanol resorcin-fuchsin and of methanol resorcin-fuchsin were compared. Formula; Dissolve 0.2 g of commercial resorcin-fuschin in 70 ml of methanol or ethanol, add 30 ml of water and 1 m1 of concentrated HC1; stain sections for 4 hours. Both solutions colored elastic and pseudoelastic fibers, cartilage and some mucins. Methanol resorcin-fuchsin also colored nuclei in methacarn- (methanol-chloroform-glacial acetic acid 6:3:1) and formalin-fixed tissues; this nuclear stain withstood counterstaining with picro-dye mictures. Zenker-fixed sections showed diffuse coloration with little or no contrast between nuclei and cytoplasm. Extraction with hot trichloracetic acid abolished binding of methylene blue, but binding of methanol resorcin-fuchsin by nuclei remained unaltered or was enhanced. Experiments with solvents containing various concentrations of methanol, ethanol or isopropanol indicated that the staining patterns of resorcin-fuchsin are determined by the nature and concentration of the alcohol. Methanol resorcin-fuchsin proved useful for simultaneous visualization of elastic tissues and nuclei.

Elastic Tissue↗
PubMed1984

The use of Light Green and Orange II as quantitative protein stains, and their combination with the Feulgen method for the simultaneous determination of protein and DNA.

The protein dyes Light Green and Orange II were studied separately and in combination with the Feulgen-Pararosanilin(SO2) and -Thionin(SO2) method for the simultaneous determination of DNA and protein. - With polyacrylamide modelfilms the pH dependency, specificity and stoichiometry of Light Green and Orange II have been investigated. The results of both staining methods with different biological objects have been compared. - In addition, the Feulgen-Thionin(SO2) method was studied with model films with respect to its specificity and stoichiometry. In biological objects it has been compared with the Feulgen-Pararosanilin(SO2) method. - When combining the Light Green staining with the Feulgen-Pararosanilin(SO2) procedure and the Orange II staining with Feulgen-Thionin(SO2), both Feulgen-DNA stainings, which were first applied, proved to be unaffected by the following protein staining procedure. When the Feulgen procedure was carried out without the dye, followed by Light Green staining, the latter became reduced when a sulfite water rinse was included but was unaffected when a running tap water rinse was used. In the case of the Orange II staining a serious reduction in dye binding capacity was found in both situations. - When the Feulgen-Pararosanilin(SO2) Light Green procedure was carried out on isolated nuclei with all dyes present, a decrease of protein dye binding was observed, similar to that found with the well-known Feulgen-Pararosanilin(SO2) Naphthol Yellow S combination. It is concluded that in spite of this reduction the latter two combinations can be used for the cytophotometric analysis of DNA and protein in the same object.

Animals↗
PubMedDate not supplied

A new device for sentinel node detection in laparoscopic colon resection.

OBJECTIVE: To test the feasibility of using a newly developed device for laparoscopic lymphatic mapping of the colon by simultaneous and quantitative detection of both tissue coloration and radioactivity. METHODS: Four pigs were used in this study. In each animal, both blue dye and radioisotope injections were utilized. Lymphatic mapping was performed laparoscopically in the sigmoid mesocolon and in the right mesocolon. A solution containing a mix of 35 microcuries of Technetium Tc-99 sulfur Colloid and 1 mL of a vital blue dye was administered subserosally by percutaneous insertion of a 25 gauge needle under laparoscopic control. The new device for automated sentinel-node detection consists of a gamma-probe coupled with a laser device and can be passed through a regular 10-mm trocar. The device detects simultaneously both radioactivity and quantitative tissue coloration. Nodes showing a radioactivity at least 5 times higher than that of the background or that had a blue colorant concentration were considered our sentinel nodes. RESULTS: Laparoscopic simultaneous and quantitative detection of sentinel nodes was feasible in all pigs. One or more sentinel nodes were identified by either the blue dye or radioisotopic technique in both the sigmoid and right mesocolon. Quantitative tissue coloration detection led to the recognition of additional nodes that were not apparently colored to the naked eye. CONCLUSION: Laparoscopic sentinel node detection using a device combining gamma and color detection is feasible in the porcine model. The significance of nodes apparently clear but positive with the quantitative detection technique should be further evaluated.

Animals↗
PubMed1988

[Use of inorganic pyrophosphatase as a marker in enzyme immunoassays].

A technique of heterogeneous enzyme immunoassay with the E. coli inorganic pyrophosphatase as marker enzyme and Malachite green dye and acidic molybdate as color reagent is developed. Color change (light-yellow/greenish blue) is extremely suitable for visual perception, in some cases making unnecessary the measuring device. Assays with pyrophosphatase are 5-10 times more sensitive than with peroxidase. Further advantages of pyrophosphatase include high thermostability, insensitivity to sodium azide, low value of Michaelis constant (5 microM), substrate stability. Examples are given of use of the pyrophosphatase for assays of human alpha-fetoprotein and immunoglobulin.

Colorimetry↗
PubMed1995

Hematoxylin staining in quantitative DNA cytometry: an image analysis study.

The suitability of commercial and pure aluminum-hematein for quantitative DNA image cytometry was investigated. Cervical smears, breast cancer aspiration biopsies, and rabbit liver tissue imprints were stained with Mayer's and Harris' al-hematein with variable staining times and dye concentrations. Pure and commercial hematoxylin was used. Nucleic acids were removed by enzyme digestion or by HCl-hydrolysis. A standard Feulgen stain served as control. DNA-polyacrylamide films were used as staining models. Absorption was measured using a VIDAS image analyzer. DNA in liver cell nuclei was not stained in a stoichiometric dye-DNA ratio. Sequential staining of cervical smears with hematein followed by the Feulgen reaction gave a covariance between 0.77 and 0.88 for IOD. Photometric errors due to unspecific RNA or protein staining were remarkable. Harris' and Mayer's hematein gave comparable results. Pure hematein gave slightly better results than commercial batches. DNA staining in model films was not quantitative with hematein. Al-hematein should therefore not be used for quantitative DNA cytometry.

Animals↗
PubMed1980

Use of a trypan blue assay to measure the deoxyribonucleic acid content and radioactive labeling of viable cells.

A simple method of determining cellular viability, DNA content, and isotopic labeling is described. Cells are sedimented onto slides and fixed in a mixture of trypan blue and paraformaldehyde which stains nonviable cells a dark blue color, whereas viable cells exclude the dye. The staining pattern is stable for at least 6 months and persists after the Feulgen reaction, allowing the selection of viable cells for cytophotometric DNA determinations. We ascertained cellular incorporation of a 3H-thymidine label by photomapping cells prior to cytophotometry and relocating these cells after preparation of autoradiographs. The method gives accurate quantitative DNA and labeling values of mouse thymocytes labeled in vitro and may be of value in studies of tumors containing a large proportion of necrotic cells.

Animals↗
PubMed1995

Chemical composition of Schiff's reagent crystals.

Crystals were prepared by adding 0.04 M sulfuric acid to a Schiff's reagent made with 2.5 g pararosaniline (PR) chloride dissolved in a saturated SO2 solution. Elemental analysis of the crystals gave the composition C19H21N3S2O7.4H2O which corresponds to the sulfate of pararosanilinesulfonic acid (PRSA) tetrahydrate. The moisture content was ca. 5%. A reagent reconstituted by dissolving 0.2 grams of crystals in HCl 0.1 N contains ca. 3.5 x 10(-3) M or 0.11% PR. A solution prepared with 2.5 g PR in 100 ml O.1 N HCl plus 0.04 M M K2S2O5 gave only a few crystals after 0.04 M sulfuric acid was added. The PR content, determined colorimetrically, was 0.25% compared with 1.35% in saturated SO2. The per cent dye loss during charcoal purification was also higher. The low concentration of PR, caused both by the lower solubility and by the larger loss during charcoal purification explains the poor yield of crystals of a reagent prepared in HCl/K2S2O5, compared to a reagent prepared in saturated SO2. After crystallization is complete, the crystals are in equilibrium with a concentration of 0.2% of PR in the supernatant: when the initial concentration is close to this value crystallization is negligible or completely fails.

Coloring Agents↗
PubMed1981

Criticism of the use of Coomassie Brilliant Blue G-250 for the quantitative determination of proteins.

The quantitative determination of proteins in biological fluids, using Coomassie Brilliant Blue G-250, was evaluated. Compared with the biuret method, the Coomassie Blue G-250 method needs a much shorter time for analysis and has a greater sensitivity. The sensitivity of the dye for albumin is significantly greater than for globulins. The standard curves for the biuret method are more linear than those for the Coomassie Brilliant Blue G-250 method. The Coomassie Brilliant Blue G-250 method produces a precipitate, which sticks to the walls of the cuvet and results in an intolerable carry-over. Therefore, the use of the Coomassie Brilliant Blue G-250 method for the quantitative determination of proteins in urine and serum is not recommended.

Chemical Phenomena↗
PubMed2002

[Sentinel lymph node mapping in colorectal cancer].

UNLABELLED: Sentinel lymph node mapping has already been accepted as part of the treatment for malignant melanomas of the skin and in breast carcinomas. The status of lymph nodes is an important prognostic marker in colorectal carcinoma as well. The authors tried the feasibility of this technique in colorectal carcinomas. The technique is analogous to the one used in breast cancer and melanoma: 2 ml of 2.5% Patentblau dye was given subserosally around the tumor. After resection the specimen was immediately sent to pathology where the lymph nodes were removed. This technique has been tried on 31 patients, 22 with colonic and 9 with rectal tumors. Of these patients, 15 were Dukes stage C, 14 were Dukes stage B and 2 were Dukes stage A. An average 4.3 blue lymph nodes were found in colon tumors and 5.4 in rectal tumors and an average 14 unstained lymph nodes were found in colon tumors, and 7 in rectal tumors. The blue nodes were predictive of the nodal status in 9 of the 15 Dukes stage C patients. In these cases the blue lymph nodes contained metastases and there were 2 cases where metastases were limited to the blue lymph nodes. SUMMARY: The authors found a high false negative rate for lymphatic mapping with the vital dye technique, therefore they try to change the method according to that used by Saha et al. The aim of sentinel node identification in colorectal carcinomas would be improved staging rather than reducing of the extent of lymphadenectomy. The role of lymphatic mapping in large bowel cancers needs further investigations. Until the results are reliable, as many lymph nodes as possible have to be excited and sent for histology.

Aged↗
PubMed2004

Effect on oximetry of dyes used for sentinel lymph node biopsy.

HYPOTHESIS: There are differences between readings of peripheral blood oxygen saturation when the effect on saturation values of methylene blue is compared with that of isosulfan blue when used in sentinel lymph node biopsy in patients with breast cancer. DESIGN: Prospective randomized study. SETTING: University tertiary care center. PATIENTS: Thirty-two women undergoing surgery for breast cancer using sentinel lymph node biopsy. INTERVENTIONS: Sentinel lymph node biopsy using methylene blue (16 patients) and isosulfan blue (16 patients); there was also a control group of 6 surgical patients in whom dyes were not used. MAIN OUTCOME MEASURES: Peripheral saturation of blood using pulse oximetry, oxygen saturation by blood-gas analysis, partial oxygen pressure by blood-gas analysis, and plasma dye levels, recorded before dye injection and 15, 30, and 60 minutes afterward. RESULTS: The 2 dyes interfered with the peripheral saturation reading, but only isosulfan blue showed significant differences. The differences in blood-gas analysis values between the 2 groups and with regard to the controls were not significant. CONCLUSIONS: Methylene blue interferes less than isosulfan blue in the peripheral saturation reading. Desaturation is factitious in both cases, and does not correspond to alterations in blood-gas analysis values.

Breast Neoplasms↗
PubMed1994

Quantitation of electrophoretically separated proteins in the submicrogram range by dye elution.

A new method for the fast, reliable and reproducible submicrogram quantitation of proteins separated by different electrophoretic techniques is presented. The method is based on a modified sensitive staining technique using Coomassie Brilliant Blue G-250 in colloidal solution combined with an optimized elution procedure of the bound dye in a 3% w/v solution of sodium dodecyl sulfate followed by photometric determination of dye concentration in the eluate. In addition a new method is provided for background correction, even suitable for gels showing strong background staining. The staining procedure allows the detection of 20 ng depending on the nature of the protein and the separation technique used. Quantitation is linear at least in the range from 50 ng to 10 micrograms and highly reproducible even under non-optimized conditions. The presented method can be applied to sodium dodecyl sulfate-electrophoresis, isoelectric focusing and two-dimensional electrophoresis.

Electrophoresis, Polyacrylamide Gel↗
PubMed1993

Gamma-probe guided localization of lymph nodes.

The initial draining lymph node (Sentinel node) of a tumour may reflect the status of the tumours spread to the remaining lymphatic bed. The sentinel node, which has been reported to predict metastatic melanoma, has recently been localized by a new invasive technique [1]. The goal of our pre-clinical trial was to test a non-invasive technique to localize the sentinel node. Gamma-probe guided localization was used to identify and then surgically remove the first draining lymph node(s) in 16 inguinal lymphatic basins of eight cats. This method was found to be comparable to an invasive method using a blue dye. Gamma-probe localization has several potential advantages in that it can: (a) precisely locate on the surface of the skin the position of an underlying lymph node, (b) intraoperatively guide the surgeon to the lymph node during dissection, (c) verify that the correct node has been biopsied, (d) determine the possible presence of residual lymph nodes, (e) allow lymph nodes to be harvested through a small incision as opposed to raising a skin flap, and (f) be rapidly and easily performed.

Animals↗
PubMed2006

[Spectrophotometric determination of chondroitin sulfate with victory blue B].

In pH 4.0 acetic acid-sodium acetate buffer solution, cationic dye victory blue B shows an absorption peak at 614 nm, and the absorption peak decreases after it reacts with chondroitin sulfate to form association particles. The decrease in absorption value is linear with chondroitin sulfate concentration in the range of 0. 1-5 microg x mL(-1), and the correlation efficient is 0.9986. The method was applied to the determination of chondroitin sulfate in synthesis samples and real samples with rapidity, simplicity and good accuracy.

Chondroitin Sulfates↗
PubMed1983

A quantitative determination of the relative amount of histones in polyacrylamide gel by silver stain.

On the basis of scanning densitometry of the stained gel, the conditions for the quantitative determination of individual histones by silver was examined and compared with the dye-staining method, in terms of higher sensitivity and faithful quantitation. Fixation with formaldehyde, coupled with simultaneous prestaining with Coomassie brilliant blue (CBB), was found to be most suitable. Prior fixation in acidic alcohol alone failed to stain the histones accurately, but this failure could be partly alleviated by prestaining with CBB. Although the sensitivity for detecting histones by silver staining is lower than that for neutral proteins by about 10-fold, it is at least 10-fold higher than the CBB stain.

Animals↗
PubMed1998

Amalgam alternatives-microleakage evaluation of clinical procedures. Part II: direct/indirect composite inlay systems.

This study investigated the degree of dye penetration of two composite inlay systems with different adhesive and bonding protocol. Forty whole extracted premolars and molars were collected. The teeth were immediately stored in water at room temperature. Class II cavity preparations were prepared and restored both, in the direct and the indirect techniques (Coltene Brilliant) and (Kulzer Estilux posterior CVS). The inlays were cemented with a composite luting cement (ART-Bond) and (Kulzer, Adhesive cement). Specimens were subjected to 750 cycles of thermal stress. They were than immersed in 2% Basic fuchsin dye. The teeth were sectioned in three planes before being ranked as to the amount of dye penetration. The data were analysed by the Wilcoxon's rank test at the 95% confidence level. There was no statistically significant difference between the direct and indirect technique considered separately for each composite inlay material, at the cervical margin location. The fuchsin staining in the occlusal area was limited to the enamel in all groups. By using the composite inlay, the factor of adaptation and bonding of composites to dentine become the significant factor contributing to leakage.

Coloring Agents↗
PubMed1983

Effects of the drug Disulphine Blue on routine biochemical investigations.

Major interference by Disulphine Blue in the colorimetric determination of amylase, albumin, protein, iron and total iron-binding capacity is described. The drug, an intense blue dye, was administered intravenously to a patient before surgery to allow demarcation of devitalised bone. Successive sampling showed the drug to have a half-life of 30 hours, to remain visible for 150 hours in both urine and blood, and to cause significant interference with the five analytes for up to 21/2 days. The 640 nm absorbance peak of Disulphine Blue was shown to be pH-dependent, and therefore the reaction conditions of individual methods may influence the degree of interference.

Blood Chemical Analysis↗
PubMed1992

Peptide mapping and microsequencing of proteins separated by SDS-PAGE after limited in situ acid hydrolysis.

A method is described for the isolation of peptide fragments from proteins separated by polyacrylamide gel electrophoresis. After completion of the electrophoresis step, gels are stained with Ponceau S or Coomassie Blue. Gel portions containing protein stained with Ponceau S are excised and transferred to borosilicate glass digestion tubes containing 0.9 ml of 1 mM NaOH or 5 mM Na2HPO4. After complete dissociation of the dye from the protein, 0.1 ml of 20% formic acid is added and the protein is hydrolyzed in situ at 112 degrees C for four hours. Subsequently the acid solution is made 10% in acetonitrile and chromatographed as such on a C18 (C4) reversed-phase column using an appropriate large-volume sample loading syringe and injection loop. Proteins stained with Coomassie Blue can be hydrolyzed in situ after complete removal of the dye with an aqueous solution containing 40% acetone, 10% triethylamine and 5% acetic acid. The gel slices are next washed with HPLC-grade water and protein is hydrolyzed in 2% formic acid under standard conditions. Gel-related contaminants do not interfere with the peptide separation under the proper conditions of HPLC analysis.

Amino Acid Sequence↗