To begin, take a slide with polymerized archived fresh tissue or paraformaldehyde fixed mouse brain tissue. Wearing eye protection, use a razor blade to separate the two glass slides of the gelling chamber. Trim the blank gel around the tissue to minimize the volume and ensure to cut asymmetrically to track the orientation of the gel after homogenization.
Gently lift the tissue containing gel from the slide with a razor blade. Transfer it into a two milliliter centrifuge tube. Fill the tube to the top with homogenization buffer preheated to 80 degrees Celsius.
Then incubate the sample with shaking at 80 degrees Celsius for eight to 60 hours. Pour the contents of the centrifuge tube into a single well of a six well plastic cell culture plate and remove the denaturation buffer with a transfer pipette. To completely remove the remaining SDS from the hydrogel, wash the sample in a 1%non-ionic surfactant solution.
Finally, store the sample in PBS and 0.02%sodium azide at four degrees Celsius.
