I have been trying to record field potentials from hippocampal slices (400um), but, not getting any response. I have tried 3 different recipies for ACSF prepartion. The slices are taken within 15 min from the time of decapitation. The slices are let to recover for 30 min before recording. I have tried stimulating the slices (10-200uA) and Mg Free ACSF with elevated KCl (4mM) for inducing epileptic activity. None of them seem to work.
1. Is there anyway to confirm that ACSF am using is working? The osmolarity is usually 308-310 and pH 7.4.
2. Is there any means of confirming tissue viablity? ( the slice looks very glittery and the edges start to fold inwards in over 2hrs)
3. I am planning to use bicuculline methiodide for inducing epilepsy in the slices. The conc i need to use is 20uM. I have 10mg vials of bicuculline. Can some tell me how to prepare stocks and how to use the drug?
Desperately waiting for an answer!!
Thanks in advance,