Material

• Dry ice (LOTS!)
• OCT embedding compound
• Microtome with good blade
• Razor blade
• Small beaker/dish with 1xPBS
• Wellplate or something pre-filled with 1xPBS or cryoprotectant to collect your slices

Steps

• Put dry ice around the center block platform. The knobs below the platform allows you to adjust the slicing stage

• While the platform (which is where you would be putting the brain) chills on dry ice, take a razor blade and cut off the cerebellum to create a flat surface so that you can rest the brain on the platform on a flat surface

• Add a small blob of OCT onto the platform

• Before the blob of OCT completely freezes, put the brain with the cut cerebellum side touching the top of the blob, and surround the brain with more dry ice

• Wait until the brain turns pale. As you slice and notice that the brain is turning back into the jelly state, add more dry ice to freeze it

• You can adjust the thickness of your slices by changing the z knob, and you can use the y-knob to move the platform in anterior-posterior direction. In the current set up, this will allow you to cut the brain into coronal slices (so cut in the dorsal-ventral direction)

• Before slicing, dab the blade with a drop of 1xPBS to wet the area with a small paintbrush

• Collect your slices with the paintbrush and make sure to re-wet the blade between each slice!