Trypan blue

TROCLOSENE POTASSIUM see POTASSIUM OICHLOROISOCYANURATE TRYPAN BLUE see C.I. DIRECT BLUE 14, TETRASOOIUM SALT... 

Fuchs VS, Golbs S, Kuhnert M, et al. 1976. [Studies into the prenatal toxic action of parathion methyl on Wistar rats and comparison with prenatal toxicity cyclophosphamide and trypan blue]. Arch Exp Vet Med 30 343-350. (German)... 

Cell count staining method with Trypan Blue (see e.g. [68])... 

Fig. 15. Effects of turbulent shear stress level and exposure time on cell viability measured by trypan blue staining. Cells were sheared in a concentric cylinder viscometer [1]...

Production of Mucosal Damage 2.3.1.2.1 Cell culture Stimulated neutrophils are known to be cytotoxic to cells in vitro (Dull et al., 1987 Dallegri et al., 1990 Grisham et al., 1990b). Several in vitro systems have been used to demonstrate oxidative damage to intestinal cells. Xanthine/XO increased Cr release and decreased [ H]thymidine uptake by IEC-18 small intestinal epithelial cell monolayers in a dose-dependent manner (Ma et al., 1991). Rat enterocytes show decreased trypan blue exclusion and increased protein release when incubated with neutrophils stimulated... 

Isolated hepatocytes incubated with ionic iron rapidly undergo lipid peroxidation. Some studies have not shown a consequent decrease in viability (as indicated by uptake of trypan blue or release of enzymes). This is probably a result of short incubation times, as changes in viability lag behind increases in lipid peroxidation, and may not occur for more than 2 h after lipid peroxidation begins (Bacon and Britton, 1990). Recent studies have shown strong correlations between increased lipid peroxidation [production of thiobarbituric acid (TBA) reactants] and loss of cell viability (trypan blue staining) (Bacon and Britton, 1989). The significance of the lag between lipid peroxidation and decreases in cell viability is as yet uncertain. 

The hypothesis is supported by two additional observations (24). First, mean cell volume for P. brevis in the absence of aponin remained constant for 8 hours, but, in the presence of aponin, a notable increase was observed within an hour and continued for eight hours. Second, Trypan blue (Cl 23850) tests indicated increased cell permeability in the presence of aponin viable, motile cells were only slightly stained swollen cells and cell debris were highly stained. 

Hwang and Bowen (2004) LNCaP Roche 10% water miscible beadlets 0.1-5.0pM DMSO in H20 6, 24, 48 h Lycopene uptake Cell viability by Trypan blue exclusion Cell cycle by flow cytometry... 

Trypan Blue exclusion and viable cell counts Cell viability by MTT... 

Trypan Blue exclusion (TB) Dead cells blue stained Cell membrane Yes3 [34]... 

Fig. 6.2. Caco-2 epithelial cell monolayers cultured with T. spiralis L1 larvae in (A) the absence or (B) presence of 1 mg ml 1 rat monoclonal, tyvelose-specific antibody 9D4 (McVay etal., 2000). Monolayers were fixed and stained with trypan blue as described in ManWarren etal. (1997). (A) Serpentine trails of nuclei in dead cells are evident, revealing the paths travelled by larvae. (B) Tyvelose-specific antibody has inhibited the migration of the larva such that it is encumbered in cell debris and has pulled up a large area of the monolayer, creating a plaque (P). Bar = 50 urn. Photomicrograph prepared by C. McVay, TTUHSC, Lubbock, Texas. 

MTX interferes with the growth of cancer cells by inhibiting the metabolism of folic acid. Drug efficacy was evaluated in vitro by MTT assay, as described above, and by Trypan Blue exclusion. Trypan Blue is a non-vital dye excluded by viable cells, but selectively staining dead cells. According to Figure 13.7, higher suppression of cell... 

Fig. 13.7 Effect of MTX-LDH hybrid on cell proliferation (A) and viability (B) as determined by MTT assay and Trypan Blue exclusion, respectively. HOS cells were incubated with free MTX, MTX-LDH hybrid or LDH for 72 h.

Lentinula edodes MnP Congo Red, Trypan Blue, Amido Black [13]... 

As the buffer used in these studies [86, 158] is hypotonic to rat peritoneal mast cells, incubations of greater than 10 min result in significant cell swelling, a large increase in membrane permeability, as judged by trypan blue staining, and eventually cell lysis. This raises some concern as to the location of the site... 

Messenger RNA molecules for both subunits of the cytochrome and the two cytosolic components are detectable in unstimulated bloodstream cells. Experiments involving incubation of neutrophil suspensions with the protein synthesis inhibitor cycloheximide indicate that constitutive expression of one or more components of the oxidase is required for the neutrophil to maintain its ability to generate reactive oxidants. For example, when neutrophils are incubated in vitro with cycloheximide, their ability to generate reactive oxidants declines more rapidly than in control cells, as they age in culture (Fig. 7.12). This decline in oxidase activity when protein biosynthesis is blocked is not due to cell death, because cells treated with cycloheximide for this time still exclude trypan blue. Furthermore, when protein biosynthesis is stimulated in neutrophils by the addition of GM-CSF for 24 h in vitro, the ability to generate reactive oxidants is enhanced considerably above the levels observed in untreated cells. 

Trypan Blue 3,3 -[(3,3 -Dimethyl[ 1,1 -biphenyl] -4,4 -diyl)bis(azo)]bis[5-amino-4-hydroxy-2,7-naphtalenedisulphonicacid]tetrasodium salt... 

The cells (1-3 x 106 cclls/rnl) were incubated for 24 h at 37 °C in RPMI1640 medium supplemented with 8mM NaHC03, 20mM HEPES, 5% FCS, 10 pg streptomycin, and lOU/ml penicillin without agents or in the presence of fullerenes C60. The number of viable cells was counted in hemocytometer using 0.4% solution of trypan blue. 

For purposes of quality control, any cell preparation used in the screen (see below) needs to have a definitive or consensus diagnosis and grading determined by histological examination. Furthermore, tumor cell preparations must be at least 80% tumor cells (as determined by cytopathological examination), whereas normal cell preparations must be devoid of any tumor cells. Cell viability, determined by trypan-blue exclusion, must be a minimum of 70%, and the signal-to-noise ratios for a predetermined cell cluster concentration must be at least threefold. 

Figure 1. Inhibition of geranylgeranoic acid-induced apoptotic cell death by a-tocophrol. Increasing concentrations (10 100 pM) of a-tocopherol were added to HuH-7 cell cultures with (closed circle) or without (open circle) 10 pM geranylgeranoic acid. After overnight incubation, the number of the viable cells was counted using a Trypan blue dye exclusion method. Means S.E. (n=3) are shown.

The antioxidant activity afforded by NO is most important if it translates into a meaningful therapeutic event. To test the importance of this reaction pathway to cell survival, we compared viability of the cells exposed to in the presence or absence of NO, When cells are exposed to 20 pM there was appreciable loss of cell membrane integrity as measured by trypan blue dye exclusion (Kelley et al, 1999) the addition of NO deaeased cell membrane damage (Figure 6). 

Figure 6. "NO protects cells from lipid peroxidation-induced cell damage. HL-60 cells at 5 x lOVml were assayed for trypan blue uptake just before iron addition in the presence or absence of NO (0.9 pM) addition, then at 7, 14, and 20 min after the initiation of peroxidation. Each data point represents the mean of at least three independent determinations and the bars the standard error. Membrane leakage was significantly different at 14 min (p=0.01) and 20 min (p=0.005). (Drawn using our data abstracted from Kelley, E.E., Wagner, B.A., Buettner, G.R., and Bums, C.P., 1999, Arch. Biochem. Biophys. 370 97-104.

Trypan blue (0.4%), a substance that is usually applied to identify dead cells, can be used as a quencher for some of the external dextran conjugates [fluorescein isothiocyanate (FITC)-dextran] (72,129). 

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