[Histonet] RE: Help with cutting mouse brain at 9-10 Microns
Smith_D <@t> kids.wustl.edu
Wed Feb 18 12:44:16 CST 2015
1. Like Pamela has asked - what's the processing schedule and what regents are used? I've had some problems if the brain is soaked in EtOH too long (70% or higher), it will cause chattering.
2. Do you slice the whole brain including cerebellum and spinal cord or did you split the hemispheres to slice them individually?
3. Since you said that you cannot get any slices more than 9-10um at all, could you describe the situation? (i.e. can't get ribbons, it was cut unevenly such as thick and thin alternatively, paraffin too hard/too soft, etc)
I mostly section rat and mouse brains at 10um without any problems. What I normally do is once the paraffin block (with brain inside of it, of course) is solidified, I tend to trim around the block around where the brain is and put it directly (the brain facing towards the ice) on the ice (NOT dry ice, just normal crushed ice) for 2-3 hours before sectioning it. After being on ice and right before sectioning, I like to rub with a small soft towel smooth the edges off around the brain then quickly dip it in the warm bath then section right away.
I do not know why I always do those above, but it sure helped me every time I slice any rodent brains. Maybe that will help you somehow!
Date: Wed, 18 Feb 2015 14:20:26 +0000
From: Kimberly Marshall <Kimberly <@t> animalreferencepathology.com<mailto:Kimberly <@t> animalreferencepathology.com>>
Subject: [Histonet] Help with cutting mouse brain at 9-10 Microns
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?Hello Histo folks
I am starting a research project with mouse brain, I am having trouble with chatter on the regular 3.5 mm sections and cant get the 9-10 mm to cut at all. I have soaked them in warm water and wonder in using a softener like conditioner would help. I am new to the animal tissue world and any advise would help.
Thanks in advance.
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