1. Prior to beginning a prep (preferably the night before), make sure you have an adequate supply of solutions and that the dissection glassware (small petri dish, small chamber, large petri dish, and vibratome chamber) is chilling in the freezer.
2. On the day of the experiment, take the ice bucket and rat transport cage downstairs. Fill the ice bucket to overflowing and retrieve animal. Put the rat cage in the hood area and set the ice bucket near the vibratome.
3. Put a beaker of 300mL external CSF into the freezer until it becomes slushy (~10 minutes)
4. For young animals, prepare a stainless steel blade (Feather S blade; FA-10; 0.1 mm) prior to beginning the dissection. The blades are packaged as attached pairs, so it is necessary to snap the blades apart. Use a single-edged razor blade to scrape the waxy coating off both sides of the vibratome blade, being careful not to touch the cutting surface of the vibratome blade. Then, clean the vibratome blade with a large cotton swab dipped in rubbing alcohol. Set the blade aside until later. For older animals you’ll use a sapphire blade. It’s normally stored in the freezer in a blue box. Do not use this without permission!
5. Fill a holding chamber with room-temperature CSF and put an oxygen line in the beaker. Place the beaker on the hot plate and heat to 30-32 degrees Celsius.
6. Get your tools set up for the dissection.Tools used include: a plastic spoon; large scissors (FST 14002-16; for decapitating the rat); scalpel (#3 holder, #10 blade; for cutting the scalp and sectioning the brain); and small angled scissors (FST 14081-09; for cutting the skull).
7. Fill the well in the vibratome (Ted Pella, Inc.; Series 1000, Pelco 101) with ice to cool the vibratome.
8. Remove your glassware and aCSF from the freezer. Press the glassware into the ice. Fill the large and small petri dishes with aCSF. The small flat dish should be placed upside down with a piece of #1 filter paper on it, this will be your cutting surface for blocking the brain. Put the beaker of remaining aCSF in the ice bucket to keep it cool.
