I blocked and mounted a brain for sectioning in a cryostat, but for some reason it didn't freeze after more than 30 minutes, despite being put at -25 C. Normally the brains I section freeze within 10 minutes. I've done nothing different with this one, but to no avail. The brain was stored in a 30% sucrose in PBS solution and was previously fixed with PFA. Any ideas?
Hi Jack,
Did you try freezing and sectioning at a lower temperature? If this doesn't work, then I would think someone might have used 10 x PBS instead of 1 x. These things do happen! You could always make up some fresh sucrose/PBS which you know to be kosher and leave it in this for a day or so (2-3 changes). If that doesn't do the trick then I'm stumped!
Best wishes
Simon
Hi Jack,
Did you try freezing and sectioning at a lower temperature? If this doesn't work, then I would think someone might have used 10 x PBS instead of 1 x. These things do happen! You could always make up some fresh sucrose/PBS which you know to be kosher and leave it in this for a day or so (2-3 changes). If that doesn't do the trick then I'm stumped!
Best wishes
Simon
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