In preparing the ACSF solution, is it a mandatory and crucial to evaluate the osmolarity of ACSF solution?
Also whether it will have influence on hippocampal slice viability?
Further, what would be the ideal incubation period in bubble chamber for the best possible slice viability.
Please, share your expertise in this regard.
Thanking you,
Best Regards,
Grandhi V Ramalingayya
It is mandatory to ascertain the osmolarity of the solution, otherwise the concentrtion may not support neuron survival. This article may help:Slice preparation Solutions ACSF: (in mM) 125 NaCl, 2.5 KCl, 2 CaCl2 ...
dendrites.esam.northwestern.edu/HippocampusDB/methods.pdf
Even if the osmolarity is not outside the range of viability for the cells it would still profoundly affect electrophysiological properties of the cells - altering resting potential and firing rates etc. I typically place the slices in acsf with oxygen in a water bath at the temperature of interest (temp also very important) and let the cells recover from the trauma of slicing for 1 hour. they typically remain viable for electrophysiology for 4 to 6 hours.
Thank you for the valuable suggestions given...
Best regards,
Grandhi V Ramalingayya
Olakunle, Your extracellular ACSF contains 25 mM of glucose, seems high than commonly used 10 mM. Any comments?
Dear Jian-young,
Thank you for the comment.
Multichannel systems (MEA) suggested to use 25 mM for ACSF and 10 mM dextrose for sucrose cutting solution.
Would you recommend 10 mM dextrose for ACSF as well??? Please let us know.
Further, what is the ideal age/ weight of the animal and also anesthesis need to be used for acute hippocampal slice preparation???
Thanking you,
Best Regards,
Grandhi V Ramalingayya