Whole mouse brain storage - (Jul/08/2010 )
I will be removing whole mouse brains to be later used for western blotting. I was planning on removing the brain, immediately flash freezing it in isopentate, then storing the frozen brain at -80 C. When I need the brain for western blotting, I plan on thawing the brain on ice or ice cold PBS, then dissect out the hippocampus. Has anyone done something similar to this before? If so, what did you do? Also, does anyone see anything wrong with my plan? Thanks for the help!
If its just the hippocampus you need why don't you just dissect out that region when you remove the brain and then freeze the required region in LN2 for -80 storage. As long as you can remove the brain and do your dissections on ice (I usually dissect the brain on the surface of a glass petri dish filled with ice to maintain the cold temp for longer) within 5-7 min, the tissue will be fine.
Well that would be the ideal situation but I don't have the time to dissect out the hippocampi from all of the mice I will be using, and we may decide to look at other brain regions along with the hippocampus at a later time. So for the sake of time, and planning for the future, it is best that I remove and save whole brains.
It should be OK, so long as you only try thawing the intact brain once! You could try OCT embedding and sectioning then using the sections to get the parts of the hippocampus by dissection. This would preserve the brain intact and mean that you could later go back and re-section to get other parts of the brain.