Evan s Blue

A dual isotope labeling technique [85] has been used to measure membrane permeability in plant cells, based on the selective permeabiHty of the membranes of living cells to tritiated water and carbon-14 labeled mannitol. Kieran [29] showed that the results of the dual isotope labeling and Evan s Blue staining methods correlated well as indicators of cell viability however, the latter was preferable in terms of reagent cost and ease of analysis. 

The effect of a particular cultivation environment on a system can be evaluated in terms of biomass (fresh/dry weight, cell number), secondary metabolite production [51,75,89,102,103,106,107] or substrate consumption (e.g. carbon source [57] or oxygen [53,108]). Using the Evan s Blue method to identify non-viable cells. Ho et al. [108] used viable cell density measurements to determine variations in specific growth rate attributable to hydrodynamic stress. 

Capillary shear tests were performed on low density (50 g fresh weight 1" ) suspensions of M. citrifolia using the apparatus illustrated in Fig. 1. Under both laminar and turbulent conditions [54,120,121], the relative viability of the suspension, evaluated using the Evan s Blue dye exclusion technique, was found to fall with exposure time in the loop. Loss of viability is well described by a first-order model ... 

Evan s Blue 6,6 -[3,3 -Dimethyl[l,l -biphenyl]-4,4 -diyl)bis(azo)bis[4-amino5-hydroxy-l,3-naphtalelenedisulphonicacid]-tetrasodium salt... 

Presumably because of the relatively high content of phenolic amino acids, products based on milk are strongly autofluorescent when viewed after either 360 nm or 490 nm wavelength excitation. Methods for fluorescence reduction such as prior staining with Toluidine Blue O or Evan s Blue can be employed, especially on sectioned material where artifactual collapse of the emulsion which is caused by the charged nature of the dye, is not an issue [22],... 

Evan s blue (Sigma), dissolved in sterile saline at 10 mg/ml... 

Muscle Fiber Integrity, Evan s Blue Staining... 

Under fluorescence (rhodamine filters), the Evan s blue fluoresces red. Healthy muscle will have positively labeled membranes, as well as blood vessels, but muscles with compromised fiber integrity will contain fibers in which the entire cross-sectioned cytoplasm is strongly fluorescent. 

Another method of Pgp regulation has been demonstrated by adrenomedullin (AM) (89). It is produced by endothelial cells in the brain and acts as a vasodilator in the cerebral circulation. It was shown that AM antisense decreased the transendo-thelial electrical resistance across endothelial monolayers. Treatment of these cells with AM activated Pgp, suggesting that AM acts as an autocrine mediator in the regulation of the properties of BBB endothelial cells. In addition, AM incubation decreased BBB permeability for sodium fluorescein (376 Da) but not for Evan s blue albumin (67 kDa). An interesting observation was that it also attenuated fluid-phase endocytosis. 

Z6. Zipf, R. E., Webber, J. M., and Grove, G. R., A comparison of routine plasma volume method using radioidinated human serum albumin and Evan s blue (T-1824) J. Lab. Clin. Med. 46, 800-805 (1955). 

To correctly position the filament, 0.2 mL of 2% Evan s Blue in saline is injected into each rat via tail vein and circulated for 30 to 60min, after which the rat is perfused transcardially with 100-mL normal saline, and the brain is removed and immediately examined. Each day, two rats of the control group and four or five rats of the drug treatment group are injected test compound dissolved in saline via tail vein at 30,60,75,120,240, or 360min after initiating occlusion to assess therapeutic efficacy. 

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