Generation of Tilted Amygdala Slices from a Mouse Brain
Transcript
Use a scalpel to cut off the cerebellum. Then, isolate the anterior portion of the brain containing the medial prefrontal cortex. Fill the cutting chamber with ice-cold cutting solution maintained at 4 degrees Celsius using a cooling unit. Oxygenate the solution.
Blot the brain dry with filter paper. Glue the 4% agar block that has been cut at a 35-degree angle to the vibratory stage, and glue the posterior part of the brain tissue onto this block. Glue two additional agar blocks in front of and behind the brain block for stability while slicing.
Place the stage with tissue attached in the cutting chamber, and ensure it is submerged. Begin cutting the acute slices. Cut each slice in half, and transfer to an interface chamber supplied with oxygenated ACSF at room temperature. After cutting the acute amygdala slices, place the interface chamber in a water bath at 36 degrees Celsius, and allow the slices to recover for 35 to 45 minutes.
