Hello. I'm currently troubleshooting an mouse aortic ring assay and I'm just trying to get the ring to respond to anything. When I add the positive control (phenylephrine in this case, although we have also tried KCL) should I just add one supraphysiological bolus? Or should I add a series of phenylephrine administrations?
Following that, when I want to add my vasorelaxant control, acetylcholine, should I wash between the phenylephrine administration? I would think this would be obvious, but I can't seem to find a solid answer on it.
Hi....here in University of Malaya, we prime the tissue with KCl; is a process like 'awakening' or activating the tissue before you start your experiment. For example, in my case, the aortic tension will be set at 1.5g. Once KCl is added, the tension rose to 2.0 g. Then, I will flush a few times till it back to 1.5g. Then PE will be added. I use single dose administration which is 10 uM. The tension will reach up to 2.0 - 2.2 g. Once it reaches the plateau, Ach will be added cumulatively starts from 0.1 uM till it reached > 50% relaxation. These steps are for endothelium integrity test. Once it has relaxed, flush out a few times until it reaches 1.5 g, then you may add PE again and test your compound of interest. Hope my explanation helped....Tq
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