I have a question regarding the health of the slices in 0 Mg2+. I have noticed that whenever I transferred adult (8 weeks) hippocampal slices to 0 Mg2+, the slices looked unhealthy within 30 min. Is that normal?
Thanks a lot.
30 mins is really fast, I think. What is your 0 Mg2+ containing? Is it the usual ACSF you use (and how long can your slices be fine in there) just without Mg2+?
I guess you are aware that 0 Mg2+ will remove the NMDAR block and thus cause massive depolarisations and Ca2+ influx in your slice (or rather into the neurons inside your slice). So it IS harmful, but I am not sure wether 30 mins is ok...
Simon is correct. Zero Mg ions will remove the block on NMDArs resulting in spontaneous bursting. Some epilepsy labs use that to model the disease.
As far as the timing, 30mins seems pretty fast, but I suspect your slices were unhealthy initially. I have recorded from healthy "looking" neurons only to witness them bursting.
If you need to do a 0 Mg experiment, patch in normal acsf, then switch out the media to 0Mg