If you use spinal cord slices, would you mind describing your procedure for cutting them?
In our lab, most people cut out the cord, and then use a block of agarose (4%) with a small half-cylinder cut out to support the cord. Then it is glued and set perpendicular to the blade on a Leica 1200VT vibratome. 300-400 um sections are then made. This works well enough but sometimes the cord pops out of the hole. Some people remove the dura, some don’t. But the ability to make the slices in this way seems to depend on the quality of this vibratome, which can cut sharply and not smash the tissue.
Some protocols I’ve seen do a full agarose embedding. This definitely helps keep the cord stable during slicing, but the fact that the cord is not surrounded by cutting solution, but only the agarose isn’t optimal for slice health.
I’m trying to use an older Leica 1000VT this summer and using the standard method from my home lab, I mostly am smashing the tissue and not making cuts.
we have another machine called the Compresstome (http://precisionary.com/) that has a different mechanism requiring full embedding. I’ve done this and macroscopically the slices are good, but the other electrophysiologists here have said the health of the slices are poor. I have yet to patch with them.
So what do you do?