I will be having freshly isolated mice brains shipped. However, these brains will be eventually used for RNA isolation and immunofluorescence staining, so it is critical that I have them delivered in a solution that will prevent RNA degradation. I have heard of RNA later but I cannot use it because it is not recommended for use with tissues that will be sectioned using a cryostat, which I will also be doing with the half of the hemisphere. Does anyone have experience/suggestions with shipping fresh mice brain for RNA isolation?
Thanks!
1- Half of a mice brain is too big to be preserved in RNA later, as the solution would not penetrate deep inside their brain to stop RNA degradation. Therefore, cut the brain in to small pieces before adding the RNA later. After all half a brain of a mice is more than enough for RNA isolation.
2- Use the other half of the mice brain for the histology. If you intend to use it for immunohistochemistry or immunohistoflourescence, then you have to preserve it in mild solution like 4% paraformaldehyde so as to prevent proteins degradation.
3- If you intend to do western blot analysis, ELISA, etc. You can still use the remaining tissues from the first half of the brain that you took samples for PCR.
Thank you.
Agreed, all the brains I harvest are sectioned half, one that is fixed in formalin and then preserve in sucrose 30% before freezing to allow for cryosectioning, and the other that goes into RNAlater for miRNA and RNA analysis. It works well but if you do dissect different sections be aware that that will completely modify your RNA profile due to different neuronal populations. Depending on what your targets are you could use only half the cortex, or only half the hypothalamus (for example) for RNAlater.