I have some mouse heart tissue embedded in OCT. Usually we take cryosections but we have decided to try and make thicker sections with a vibratome. I was wondering if anyone has experience making vibratome sections from OCT embedded tissue. Or would it be better to re-embed the tissue in agarose and then do section?
I am grateful for any help you can provide.
Pieter
Dear Pieter,
Is the tissue fixed and cryoprotected, or was it flash frozen in OCT? Either way, you will probably need to thaw the tissue to get rid of the OCT. It needs to be frozen to remain solid, and I don't know of a vibratome that gets THAT cold.
Does someone in your institution have a sliding microtome with a freezing stage (holds the dry ice)? That would be a much easier way to cut thick frozen sections (30-50 um). There are also a few cryostats models that will let you cut thicker sections. Sometimes the greater section thickness settings are under the "trim" feature.
Good Luck!
Jill
Dear Pieter,
Is the tissue fixed and cryoprotected, or was it flash frozen in OCT? Either way, you will probably need to thaw the tissue to get rid of the OCT. It needs to be frozen to remain solid, and I don't know of a vibratome that gets THAT cold.
Does someone in your institution have a sliding microtome with a freezing stage (holds the dry ice)? That would be a much easier way to cut thick frozen sections (30-50 um). There are also a few cryostats models that will let you cut thicker sections. Sometimes the greater section thickness settings are under the "trim" feature.
Good Luck!
Jill
Thank you both. I agree with you both of you that vibratome sections of OCT embedded tissue won't work so I am not gonna try it out. For now I will continue by just thawing the OCT and re-embedding the tissue in agarose for sections. This seems to work just fine.
Thanks again!
Pieter
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