I am looking to cut a mouse in half along the spine to insert a piece of film between both sides. Literature for whole body fixation suggests transcardial perfusion, but I need to keep the body intact (as much as possible) and TC perfusion is very invasive. Immersion fixation does not seem like a good choice considering the amount of time it takes to permeate the tissue, and the size of a mouse. Alternatively, I could freeze the animal (LiN2? -20C freezer?) and then somehow cut it? Any suggestions would be appreciated!
Note: I am not particularly concerned about maintaining tissue integrity for pristine histology purposes, but want to minimize any tissue lost due to cutting and need to keep the spinal cord as intact as possible (yes, nearly impossible, I know).
Thanks!
Hello,
I suppose it is not so difficult to cut a dead mouse in the medio sagittal axis ventrally (thorax and abdomen) to perfuse her transcardially and finally cut her into two pieces. However, I think the bone of the spine will not be easily be cutted and so the spinal will be more or less crushed.
Hope this helps, kind regards
Stefan
The difficult thing about the subject is the process of cutting the spine and can be overcome through use of formic or nitric acid with the fixative to decalcify the spine ...try may be useful
with my regards
In a follow up to my original question, I have fixed some whole mice in paraformaldehyde. Now that we are ready to move forward with cutting down the spine (using a saw), I am wondering what the best material would be for embedding. Paraffin? A stiff Agar or Gelatin? Something like a Sylgard elastomer kit? I imagine the blade will heat up some, so perhaps a high melting point wax would be best? Many thanks for your ideas!
Hey folks!
In case anyone is interested in trying this themselves, we had great results by first fixing the mice using whole-body submersion in 4% PFA for > 1 week (with small cut to abdomen to facilitate access to gut etc), followed by embedding in Gulf wax (paraffin wax - from the grocery store) or in Sylgard 164 (1:10 ratio) and letting set for 3 days. We imaged both blocks with CT to identify the location of the spinal cord, and then cut along the axis (actually, slightly to the right of the line) using a Grizzy saw (blade diameter ~ 0.5mm). Both mice cut very well (paraffin being the easiest) and we were able to produce mouse halves which both had well preserved spinal cord tissue (see photo: left = Elastomer, right = Wax). Thanks for all the feedback and suggestions.
Janet
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