I am trying to cut mouse brain fixed in 10% formalin for 24 hours embedded in OCT after sucrose treatment. The samples sink in the sucrose, so we don't think that is the problem and the tissue looks completely fixed based on color, firmness, etc. The issue is that our samples splinter when cut. I have heard this is because the sample is too cold, but we have tried at a range of temperatures and the problem persists. I have also tried adjusting blade angle and changing the blade, but this did not seem to help.
Does anyone have any suggestions or a good protocol to follow?
Thanks,
Usually, when the tissue is overly brittle/hard, splinters may occur. Even though fixed tissues are more stable and resistant to degradation, it will increase the hardness of the tissues.
You can try reducing the formalin conc. or fixation time if the fixation step is mandatory for your application.
Otherwise try a fresh tissue-cryosectiong (without fixation), if possible.
Please keep in mind that, the brain tissues are very fragile and heterogeneous and are usually difficult for cryosectioning. If your application permits routine paraffin-embedded sectioning, you can preserve a good morphology.
Thanks.
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