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At least 199 records · Page 11Linked to original sources
PubMed2003

Trypan blue staining of internal limiting membrane and epiretinal membrane during vitrectomy: visual results and histopathological findings.

AIMS: To report on the use of trypan blue (TB) 0.06% for staining the internal limiting membrane (ILM) and epiretinal membrane (ERM) during vitrectomy and report on their histology. METHOD: 14 consecutive patients with idiopathic macular hole or macular pucker (seven patients each) were prospectively recruited for ILM or ERM peel respectively. After pars plana vitrectomy and induction of posterior vitreous detachment, 0.5 ml TB 0.06% in phosphate buffered saline (VisonBlue) was injected over the posterior pole in an air filled eye and left for 2 minutes. The stained tissue was peeled with intraocular forceps. Specimens were evaluated using histochemical and immunohistochemical methods. RESULTS: The average follow up was 4.4 months. Internal limiting membranes and epiretinal membranes were stained satisfactorily in all cases and removed successfully. Eight patients (57%) had improvement of 2 or more Snellen lines. All seven macular holes closed. In the ERM cases, no residual membranes were observed clinically, at the latest follow up. No complications relating to the use of the dye were encountered intraoperatively or postoperatively. Of the 14 procedures, nine (four macular hole and five macular pucker) yielded sufficient tissue for histopathological evaluation. Histological and immunohistological assessment revealed that the morphology of these specimens was similar to that observed in macular hole ILM and macular pucker ERM removed without the aid of dye. CONCLUSION: TB staining facilitated the identification and delineation of ILM and ERM removal during the surgical management of macular holes and macular pucker. The visual outcome of this series and the specimens removed suggest they are no different from those without TB staining. Its use in posterior segment appears to be safe but further studies are required to investigate its long term safety.

Adult↗
PubMed1975

Effect of morphine, hydromorphone, methadone, mescaline, trypan blue, vitamin A, sodium salicylate, and caffeine on the serum interferon level in response to viral infection.

The ability of morphine, hydromorphone, methadone, mescaline, trypan blue, vitamin A, sodium salicylate and caffeine to alter the circulating serum level of interferon was investigated in mice. Sodium salicylate, and caffeine increased the serum level of interferon; however, the interferon increase produced by sodium salicylate was dose-dependent, i.e. low doses increased interferon titers, high doses decreased the titers. All other compounds decreased the circulating level of interferon. These results suggest that compounds of varied structure and biological activity, and possessing teratogenic potential, are capable of depressing the circulating level of interferon. Some of these "anti-interferon" compounds, i.e. morphine, hydromorphone, methadone, depressed interferon levels at all doses whereas sodium salicylate decreased the interferon level only at relatively high dose levels.

Animals↗
PubMed2004

Trypan blue- and indocyanine green-assisted epiretinal membrane surgery: clinical and histopathological studies.

PURPOSE: To evaluate the clinical outcome and electron microscopic findings of trypan blue (Tb) and indocyanine green (ICG) assisted epiretinal membrane (ERM) surgery. METHODS: This is a prospective consecutive noncomparative interventional case series. After pars plana vitrectomy, 0.1 ml of 0.6 mg/ml Tb solution was applied for 1 min under air for ERM staining. After ERM removal, internal limiting membrane (ILM) was further peeled after staining with 0.2 ml of 1 mg/ml ICG solution. Intraoperative specimens were sent for electron microscopy. Tb was considered useful if the edge of ERM was stained where peeling could be initiated with a clearer visualisation of the overall extent of the ERM. RESULTS: In all, 16 eyes from 16 patients were recruited. There were nine grade 1 ERMs, five grade 2 ERMs, and two grade 3 ERMs. Tb was useful in six (67%) of the nine eyes with grade 1 ERMs and in all eyes with grade 2 or 3 ERMs. The three remaining grade 1 ERMs were removed together with surrounding ILM that was stained by ICG. The mean line of improvement was 1.3 lines with the median BCVA improved from 6/12 to 6/9. All 16 eyes had symptomatic improvement and none developed ERM recurrence. No complication related to Tb or ICG was observed clinically or angiographically. Electron microscopy of the Tb-stained ERM specimens showed fragments of ILM in all specimens. CONCLUSIONS: Tb and ICG are useful intraoperatively to improve the visualisation and facilitate complete removal of ERM and ILM in macular ERM surgery.

Aged↗
PubMed1981

Dual staining of corneal endothelium with trypan blue and alizarin red S: importance of pH for the dye-lake reaction.

Evaluation of corneal endothelial integrity by combined staining with the vital stain trypan blue and the intercellular stain alizarin red S provides a simple, quick technique for visualisation of both damaged and normal cells, thereby permitting the quantification of endothelial cell damage. Adjustment of the pH of the alizarin red S reagent to 4.2 is important for optimum dye-laking at the intercellular borders, and brief fixation with glutaraldehyde maintains the staining effect of both dyes.

Animals↗
PubMed2005

Photodynamic actions of indocyanine green and trypan blue on human lens epithelial cells in vitro.

PURPOSE: The purpose of this study was to evaluate the toxicity and photodynamic activity of indocyanine green (ICG) and trypan blue (TryB) on cultured human lensepithelial cells (LECs). DESIGN: Experimental study. METHODS: Lens epithelial cell viability was assessed after treatment with ICG and TryB concentrations ranging from 0.025 to 5.0 mg/ml, and exposure to 806 nm diode laser. RESULTS: At ICG concentrations below 0.5 mg/ml, there was > or =75% cell viability; at higher ICG concentrations there was dose-dependent cytotoxicity in addition to loss of cellular viability due to ICG photosensitization. TryB had little cytotoxicity to the LECs: >80% cells were viable irrespective of the dye concentration or laser treatment. CONCLUSIONS: These data indicate that ICG may have application as a photosensitizer in the selective eradication of residual LECs after cataract surgery to reduce the incidence of posterior capsule opacification.

Cell Count↗
PubMedDate not supplied

Trypan blue staining of the anterior capsule under an air bubble with a modified cannula.

To attain good visibility of the anterior capsule in the advanced or white cataract, trypan blue 0.1% is used to stain the anterior capsule. The dye is usually injected under an air bubble. However, it is difficult to inject the dye properly due to capillary forces. An ordinary anterior chamber cannula was modified and its coverage area increased to facilitate the staining of the anterior capsule under an air bubble. The anterior capsule was stained properly by using the modified cannula in all of the cases.

Aged↗
PubMed1997

Simultaneous in situ detection of apoptosis and necrosis in monolayer cultures by TUNEL and trypan blue staining.

A method for simultaneously detecting membrane permeability (characteristic of necrosis) and DNA fragmentation (characteristic of apoptosis) is described. By combining a common dye-exclusion method (Trypan Blue) with a commercially available terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) labeling kit, we have succeeded in developing a novel methodology for obtaining permanently mounted slides of monolayer cell cultures double-labeled for DNA fragmentation and cell lysis. This method should facilitate in situ studies of cell death by allowing for a more accurate quantification of total toxicity in monolayer cell cultures and perhaps further enhance our understanding of the different mechanisms of cell death as well.

Apoptosis↗
PubMed2005

A comparison of outcomes after indocyanine green and trypan blue assisted internal limiting membrane peeling during macular hole surgery.

AIMS: To compare the anatomical and visual outcomes of macular hole repair surgery using indocyanine green (ICG) or trypan blue (TB) staining of the internal limiting membrane (ILM). METHOD: Retrospective analysis of 37 eyes from 37 consecutive patients with stage 2, 3, and 4 idiopathic macular holes who underwent macular hole repair by one surgeon using the same technique but utilising different dyes for ILM peeling. In 19 patients ICG was used while 18 patients had TB. The anatomical and visual results in these two groups were compared. RESULTS: There were no significant differences in the demographic and macular hole characteristics of the ICG and TB groups. Macular hole closure was achieved in 91.9% of all patients of which the ICG group had an 89.5% hole closure rate and the TB group had a 94.4% hole closure rate. After excluding cases with failed hole closure and other vision affecting complications, there was no significant difference between the preoperative visual acuities in the TB and ICG groups but the postoperative visual acuities were better in the TB than the ICG group (p = 0.036). The TB group also had more Snellen lines of improvement than the ICG group (2.94 v 1.79 lines; p = 0.046). CONCLUSION: TB appears to be less toxic than ICG when used in dye assisted peeling of ILM during macular hole repair as reflected by the better visual results in the TB group of patients.

Aged↗
PubMed2006

Hypothermia of 8 degrees C protects cultured retinal pigment epithelial cells and retinal ganglion cells against trypan blue toxicity.

PURPOSE: To determine whether hypothermia of 8 degrees C can protect cultured human retinal pigment epithelial (ARPE-19) cells and rat retinal ganglion cells (RGC-5) against trypan blue (TB) toxicity. DESIGN: Laboratory investigation. METHODS: ARPE-19 cells and RGC-5 were exposed to balanced salt solution as controls, and 0.05% and 0.5% TB at 37 degrees C, and at 8 degrees C for one minute. The percentage of surviving cells was determined by the resazurin test. RESULTS: TB induced a statistically significant decrease in the percentage of ARPE-19 cells surviving at 0.5% TB at 37 degrees C (P < .01). Conversely, TB induced a statistically significant decrease in the percentage of RGC-5 surviving at all conditions except for 0.05% TB at 8 degrees C (0.05% 37 degrees C; P < .05, 0.5% 37 degrees C and 8 degrees C; P < .01). CONCLUSIONS: These results indicate that reducing the temperature to 8 degrees C has a protective effect against the TB toxicity for ARPE-19 cells and RGC-5 in culture.

Animals↗
PubMed1981

Trypan blue dye uptake and lactate dehydrogenase in adult rat hepatocytes--freshly isolated cells, cell suspensions, and primary monolayer cultures.

Leakage of lactate dehydrogenase and staining by the vital dye trypan blue were investigated in adult rat hepatocytes at the time of isolation, in suspensions up to 3 h and in primary monolayer cultures up to 3 d. These two parameters of plasma membrane integrity were found to correlate closely in hepatocyte suspensions, but to a lesser degree in monolayer cultures. Functional activity was demonstrated in culture by glucose consumption and lactic acid production. There was a balance of total lactate dehydrogenase (LDH) activity over time for both hepatocyte suspensions and cultures. Loss of LDH activity in the cell fraction was accompanied by a corresponding increase in enzyme activity in the media fraction. Lactate dehydrogenase activity per dye-excluding hepatocyte was calculated to be 9.2 +/- 1.5 X 10(-6) IU assayed at 37 degrees C for 25 preparations of isolated hepatocytes. The results suggest that leakage of cytoplasmic enzyme and vital dye staining are of comparable sensitivity in evaluating hepatocyte preparations. Measurement of LDH leakage offers a less subjective alternative to cell counting procedures and is applicable to both attached and suspended cells.

Animals↗
PubMed1977

Fluorescence of Trypan blue in frozen-dried embryos of the rat.

Freeze-drying and fluorescence microscopy techniques were combined to create a sensitive method for the visualization of the teratogenic dye, Trypan blue, in both protein-bound and free forms. In the development and initial application of this method, visceral yolk sacs of several gestational ages as well as normal appearing, 12-day embryos obtained from dye-injected rats were utilized. Observations on paraffinized sections of the yolk sac placentae demonstrated that only the protein-bound form of the dye exists in the yolk sac cavity whereas both forms of the dye exist in supranuclear regions of cells of the visceral endoderm. Paraffin sections of the normal appearing, 12-day embryos displayed the protein-bound form of dye within lumina of mid- and hind-gut, and both forms of dye in the primitive mucosa of mid- and hind-gut. The advantages of the method are derived not only from the use of fluorescence microscopy but also from the avoidance of solvents that are employed in more routine microtechniques.

Animals↗
PubMed1978

Azo reduction of trypan blue to a known carcinogen by a cell-free extract of a human intestinal anaerobe.

The azo reductase activity of a cell-free extract of Fusobacterium sp. 2 is characterized using trypan blue as a substrate. Either chemical reduction of this dye with sodium hydrosulfite or reduction by the cell-free extract produces a mutagenic product, o-tolidine. The o-tolidine is mutagenic in the Ames Salmonella/mammalian-microsome mutagenicity test when activated by a rat liver S9 preparation.

Benzidines↗
PubMed2000

Inhibition of formation of protease-resistant prion protein by Trypan Blue, Sirius Red and other Congo Red analogs.

Five compounds related to Congo Red were found to inhibit generation of protease-resistant prion protein in a cell-free system. In this assay Trypan Blue, Evans Blue, Sirius Red F3B, Primuline and Thioflavin-S were all more inhibitory than Congo Red itself. In scrapie-infected mouse neuroblastoma cells one compound, Sirius Red F3B, was capable of blocking the formation of protease-resistant prion protein to a similar extent as Congo Red; however, the other four compounds were less effective. Some of these compounds should be considered for testing in TSE disease models in live animals.

Animals↗
PubMed1978

The use of trypan blue for counterstaining in the Sepharose bead immunofluorescence test. The application of the test for the demonstration of primate retrovirus-specific antibodies and antigens.

The Sepharose bead immunoflurorescence test was performed by counterstaining the beads with trypan blue. This results in a red staining of negative beads which allows an easy distinction from positive green-fluorescent beads. Sepharose beads conjugated with viral proteins or antiviral antibodies were used to demonstrate Mason-Pfizer monkey virus (MPMW)- and simian sarcoma virus (SSV) - specific antigens or antibodies. The test shows a high sensitivity and specificity and needs a small amount of material.

Antibodies, Viral↗
PubMed1986

Pasteurella haemolytica leukotoxin: comparison of 51chromium-release, trypan blue dye exclusion, and luminol-dependent chemiluminescence-inhibition assays for sensitivity in detecting leukotoxin activity.

Dilutions of concentrated, dialyzed Pasteurella haemolytica culture supernatant were caused to react with bovine neutrophil (PMN) suspensions, and then the trypan blue dye exclusion (TBDE), 51chromium (51Cr)-release, and luminol-dependent chemiluminescence-inhibition (LDCLI) assays were done to compare their relative sensitivities in detecting biological activity of P haemolytica leukotoxin (cytotoxin). The culture supernatant was concentrated approximately 200:1, and when caused to react as an undiluted preparation with bovine PMN, it was cytotoxic for 38.6% and 80.4% of PMN as determined by TBDE and 51Cr-release assays, respectively. This undiluted leukotoxin preparation caused 100% inhibition of the luminol-dependent chemiluminescence responses of bovine PMN. The LDCLI assay was the most sensitive of the 3 in vitro assays for P haemolytica leukotoxin activity--being approximately 17 times and 2,480 times more sensitive than the 51Cr-release and TBDE assays, respectively. The relative advantages and disadvantages of the 3 assays as in vitro systems for detecting and titrating leukotoxin activity and investigating the role of leukotoxin in disease pathogenesis and immunity are discussed. Because of its sensitivity, specificity, economy, technical ease, and potential for adaptation to automation, the LDCLI assay would seem to be the in vitro assay of choice for quantitating P haemolytica leukotoxin activity. To aid standardization of studies of leukotoxin between different laboratories, it is suggested that P haemolytica leukotoxin be quantitated and expressed as chemiluminescence inhibitory units.(ABSTRACT TRUNCATED AT 250 WORDS)

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↗
PubMed2003

Double vital staining using trypan blue and infracyanine green in macular pucker surgery.

AIMS: To study the clinical properties of double vital staining in premacular fibrosis, facilitating complete removal of all epiretinal tissue. METHODS: In a two step surgery, the epiretinal pucker was removed after staining with trypan blue, whereafter the inner limiting membrane was peeled after staining with infracyanine green. RESULTS: In all 30 patients, a separate epiretinal layer and inner limiting membrane were removed from the macular area. Pathological examination showed different histological properties of the removed layers. An increased visual acuity was measured in 26 patients, and a slightly decreased visual acuity in one patient. CONCLUSION: The described double staining technique could be a novel valuable tool that may help to achieve optimal anatomical and functional recovery after surgery for premacular fibrosis

Aged↗