Is it possible to identify the stress fiber formation using phalloidin conjugated fluorescent dye ?
phalloidin conjugated fluoreecent dyes stain all actin fillaments, therefore, it might be suitable if you co-stain actin with myosin II, and if your cells are fibroblasts, it would be a good idea to stain alpha-smooth muscle actin, since this protein is recruited to the stress fibers
Might there be a means of measuring interfacial water stress (IWS) in vitro, or invivo (intravitally or inter vivos) as a biomarker of inflammation? Might nutritional stress generate IWS? Might nutritional stress be a sufficient cause of IWS? It's NOT solely the proteins that are being stained. It's the solvated proteins that are stained, yes? The interphase, including solutes, HSPGs, and vicinal water in the glycocalyces, are probably also being stained, yes?
I meant that after phalloidin-fluorescent staining is it possible to differentiate by observing the image/ by using a phase contrast / confocal microscope. Is there any difference in the pattern or staining property ?
Sorry for my naivete! Is this an intravital technique? Are you aware of any intravital microscopic measures of inflammation, generally? Gallez and Coakley noted interfacial "waves" on the cell membranes of various cell types by light microscopy, intravitally, e.g. RBCs, platelets, monocytes, endothelial cells. This should be systematically explored. The potential clinical significance is considerable.
absolutely you can see clearly the difference between stressed cells and non stressed ones by the mentioned fluorescence stain and using the most basic fluorescence microscope. the F actin stress fibers, bundles, should be obvious in normal proliferating cells, (the non stressed ones) as in stressed cells (usually serum starved) you have the Rho proteins at the baseline level.
I recommend Phalloidin-Atto 647N (Sigma) that stains actin filamnets in cells after staining shortly with 70% Ethanol (2min), then 2xPBS washing, folowed by incubation with Phalloidin (1:1000) for 1 hour in darkness RT, 2xPBS washing, then the cells are embeded in VectaShield and I observe my cells in confocal microscope. The Phalloidin emmision is in infra red and gives bright, strong signal.
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